Inhibitors of akt activity

ABSTRACT

The instant invention provides for substituted naphthyridine compounds that inhibit Akt activity. In particular, the compounds disclosed selectively inhibit one or two of the Akt isoforms, preferably Akt1. The invention also provides for compositions comprising such inhibitory compounds and methods of inhibiting Akt activity especially Akt1 by administering the compound to a patient in need of treatment of cancer.

BACKGROUND OF THE INVENTION

The present invention relates to substituted naphthyridine compoundswhich are inhibitors of the activity of one or more of the isoforms ofthe serine/threonine kinase, Akt (also known as PKB; hereinafterreferred to as “Akt”). The present invention also relates topharmaceutical compositions comprising such compounds and methods ofusing the instant compounds in the treatment of cancer.

Apoptosis (programmed cell death) plays essential roles in embryonicdevelopment and pathogenesis of various diseases, such as degenerativeneuronal diseases, cardiovascular diseases and cancer. Recent work hasled to the identification of various pro- and anti-apoptotic geneproducts that are involved in the regulation or execution of programmedcell death. Expression of anti-apoptotic genes, such as Bcl2 or Bcl-xL,inhibits apoptotic cell death induced by various stimuli. On the otherhand, expression of pro-apoptotic genes, such as Bax or Bad, leads toprogrammed cell death (Adams et al. Science, 281:1322-1326 (1998)). Theexecution of programmed cell death is mediated by caspase-1 relatedproteinases, including caspase-3, caspase-7, caspase-8 and caspase-9 etc(Thornberry et al. Science, 281:1312-1316 (1998)).

The phosphatidylinositol 3′-OH kinase (PI3K)/Akt pathway appearsimportant for regulating cell survival/cell death (Kulik et al. Mol.Cell. Biol. 17:1595-1606 (1997); Franke et al, Cell, 88:435-437 (1997);Kauffmann-Zeh et al. Nature 385:544-548 (1997) Hemmings Science,275:628-630 (1997); Dudek et al., Science, 275:661-665 (1997)). Survivalfactors, such as platelet derived growth factor (PDGF), nerve growthfactor (NGF) and insulin-like growth factor-1 (IGF-1), promote cellsurvival under various conditions by inducing the activity of PI3K(Kulik et al. 1997, Hemmings 1997). Activated PI3K leads to theproduction of phosphatidylinositol (3,4,5)-triphosphate(PtdIns(3,4,5)-P3), which in turn binds to, and promotes the activationof, the serine/threonine kinase Akt, which contains a pleckstrinhomology (PH)-domain (Franke et al Cell, 81:727-736 (1995); HemmingsScience, 277:534 (1997); Downward, Curr. Opin. Cell Biol. 10:262-267(1998), Alessi et al., EMBO J. 15: 6541-6551 (1996)). Specificinhibitors of PI3K or dominant negative Akt mutants abolishsurvival-promoting activities of these growth factors or cytokines. Ithas been previously disclosed that inhibitors of PI3K (LY294002 orwortmannin) blocked the activation of Akt by upstream kinases. Inaddition, introduction of constitutively active PI3K or Akt mutantspromotes cell survival under conditions in which cells normally undergoapoptotic cell death (Kulik et al. 1997, Dudek et al. 1997).

Three members of the Akt subfamily of second-messenger regulatedserine/threonine protein kinases have been identified and termedAkt1/PKBα, Akt2/PKBβ, and Akt3/PKBγ (hereinafter referred to as “Akt1”,“Akt2” and “Akt3”), respectively. The isoforms are homologous,particularly in regions encoding the catalytic domains. Akts areactivated by phosphorylation events occurring in response to PI3Ksignaling. PI3K phosphorylates membrane inositol phospholipids,generating the second messengers phosphatidyl-inositol3,4,5-trisphosphate and phosphatidylinositol 3,4-bisphosphate, whichhave been shown to bind to the PH domain of Akt. The current model ofAkt activation proposes recruitment of the enzyme to the membrane by3′-phosphorylated phosphoinositides, where phosphorylation of theregulatory sites of Akt by the upstream kinases occurs (B. A. Hemmings,Science 275:628-630 (1997); B. A. Hemmings, Science 276:534 (1997); J.Downward, Science 279:673-674 (1998)).

Phosphorylation of Akt1 occurs on two regulatory sites, Thr³⁰⁸ in thecatalytic domain activation loop and on Ser⁴⁷³ near the carboxy terminus(D. R. Alessi et al. EMBO J. 15:6541-6551 (1996) and R. Meier et al. J.Biol. Chem. 272:30491-30497 (1997)). Equivalent regulatoryphosphorylation sites occur in Akt2 and Akt3. The upstream kinase, whichphosphorylates Akt at the activation loop site has been cloned andtermed 3′-phosphoinositide dependent protein kinase 1 (PDK1). PDK1phosphorylates not only Akt, but also p70 ribosomal S6 kinase, p90RSK,serum and glucocorticoid-regulated kinase (SGK), and protein kinase C.The upstream kinase phosphorylating the regulatory site of Akt near thecarboxy terminus has not been identified yet, but recent reports provideevidences indicating that following molecules mediate this event; therapamycin insensitive mammalian target of rapamycin complex (mTORC2) (D.D. Sarbassov et al. Science 307: 1098-1101 (2007)), integrin-linkedkinase (ILK-1) (S. Persad et al. J. Biol. Chem. 276: 27462-27469(2001)), PDK1 (A. Balendran et al. Curr Biol. 9: 393-404 (1999)),DNA-dependent protein kinase (DNA-PK) (J Feng et al. J. Biol. Chem. 279:41189-41196 (2004)), and AKT itself (A. Toker and A. C. Newton. J. Biol.Chem. 275: 8271-8274 (2000)).

Analysis of Akt levels in human tumors showed that Akt2 is overexpressedin a significant number of ovarian (J. Q. Cheng et al. Proc. Natl. Acad.Sci. U.S.A. 89:9267-9271 (1992)) and pancreatic cancers (J. Q. Cheng etal. Proc. Natl. Acad. Sci. U.S.A. 93:3636-3641 (1996)). Similarly, Akt3was found to be overexpressed in breast and prostate cancer cell lines(Nakatani et al. J. Biol. Chem. 274:21528-21532 (1999).

The tumor suppressor PTEN, a protein and lipid phosphatase thatspecifically removes the 3′ phosphate of PtdIns(3,4,5)—P3, is a negativeregulator of the PI3K/Akt pathway (Li et al. Science 275:1943-1947(1997), Stambolic et al. Cell 95:29-39 (1998), Sun et al. Proc. Natl.Acad. Sci. U.S.A. 96:6199-6204 (1999)). Germline mutations of PTEN areresponsible for human cancer syndromes such as Cowden disease (Liaw etal. Nature Genetics 16:64-67 (1997)). PTEN is deleted in a largepercentage of human tumors and tumor cell lines without functional PTENshow elevated levels of activated Akt (Li et al. supra, Guldberg et al.Cancer Research 57:3660-3663 (1997), Risinger et al. Cancer Research57:4736-4738 (1997)).

These observations demonstrate that the PI3K/Akt pathway plays importantroles for regulating cell survival or apoptosis in tumorigenesis.

Inhibition of Akt activation and activity can be achieved by inhibitingPI3K with inhibitors such as LY294002 and wortmannin. However, PI3Kinhibition has the potential to indiscriminately affect not just allthree Akt isozymes but also other PH domain-containing signalingmolecules that are dependent on PdtIns(3,4,5)—P3, such as the Tec familyof tyrosine kinases. Furthermore, it has been disclosed that Akt can beactivated by growth signals that are independent of PI3K.

Alternatively, Akt activity can be inhibited by blocking the activity ofthe upstream kinase PDK1. No specific PDK1 inhibitors have beendisclosed. Again, inhibition of PDK1 would result in inhibition ofmultiple protein kinases whose activities depend on PDK1, such asatypical PKC isoforms, SGK, and 56 kinases (Williams et al. Curr. Biol.10:439-448 (2000).

It was also reported that deficiency of Akt1 is sufficient to inhibittumorigenesis in several genetically modified mice tumor models, such asPTEN+/−model as prostate tumors (M. L. Chen et al. Genes & Dev.20:1569-1574 (2006)), MMTV-ErbB2/Nue and MMTV-polyoma middle Ttransgenic mice as breast tumor models (I. G. Maroulakou et al. CancerRes. 67: 167-177 (2007)).

Mice lacking the Akt2 showed diabetes mellitus-like syndrome by theimpairment in the ability of insulin to lower blood glucose (J. L.Thorvaldsen et al. Science. 292: 1728-1731 (2001) and R. S. Garofalo etal. J. Clin. Invest. 112:197-208 (2003)).

The compounds of the instant invention have unexpected advantageousproperties over the cyclopropyl substituted naphthyridine compoundsspecifically described in WO 2006/135627.

It is an object of the instant invention to provide novel compounds thatare inhibitors of Akt, especially Akt1.

It is also an object of the present invention to provide pharmaceuticalcompositions that comprise the novel compounds that are inhibitors ofAkt, especially Akt1.

It is also an object of the present invention to provide a method fortreating cancer that comprises administering such inhibitors of Aktactivity, especially Akt1.

SUMMARY OF THE INVENTION

The instant invention provides for substituted naphthyridine compoundsthat inhibit Akt activity. In particular, the compounds disclosedselectively inhibit one or two of the Akt isoforms, preferably Akt1. Theinvention also provides for compositions comprising such inhibitorycompounds and methods of inhibiting Akt activity, especially Akt1 byadministering the compound to a patient in need of treatment of cancer.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the instant invention are useful in the inhibition ofthe activity of the serine/threonine kinase Akt, especially Akt1. In afirst embodiment of this invention, the inhibitors of Akt activity areillustrated by the Formula A:

wherein:

E, F, G, H, I, J, K, L and M are independently selected from: C or N,wherein each E, F, G, H, I, J, K, L and M is optionally substituted withR¹;

a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; p is independently 0, 1, 2, 3,4 or 5;

Ring Y is (C₄-C₇)cycloalkyl, said cycloalkyl is substituted with R^(x),and further optionally substituted with one or more substituentsselected from: (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, CO₂H,halo, CN, OH and NR⁷R⁸, said alkyl, cycloalkyl and alkoxy is optionallysubstituted with one or more substituents selected from halo, CN, OH andNR⁷R⁸;

Ring Z is selected from: (C₃-C₈)cycloalkyl, aryl, heteroaryl andheterocyclyl;

R¹ is selected from: H, oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)_(m)NR⁷R⁸, SH,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶;

R² is independently selected from: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, SH, S(O)_(m)NR⁷R⁸,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶;

R⁶ is: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo, CN, OH,O_(b)C₁-C₆ perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO, (N═O)R⁷R⁸,S(O)_(m)NR⁷R⁸, SH, S(O)_(m)—(C₁-C₁₀)alkyl or(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, said alkyl, aryl, alkenyl, alkynyl,heterocyclyl, and cycloalkyl optionally substituted with one or moresubstituents selected from R^(6a);

R^(6a) is selected from: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,O_(a)(C₁-C₃)perfluoroalkyl, (C₀-C₆)alkylene-S(O)_(m)R^(a), SH, oxo, OH,halo, CN, (C₂-C₁₀)alkenyl, (C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl,(C₀-C₆)alkylene-aryl, (C₀-C₆)alkylene-heterocyclyl,(C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a), (C₀-C₆)alkylene-CO₂R^(a), C(O)H,and (C₀-C₆)alkylene-CO₂H, said alkyl, alkenyl, alkynyl, cycloalkyl,aryl, and heterocyclyl is optionally substituted with up to threesubstituents selected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, and N(R^(b))₂;

R⁷ and R⁸ are independently selected from: H,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)-heterocyclyl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, SH, SO₂R^(a), and (C═O)_(a)NR^(b) ₂, said alkyl,cycloalkyl, aryl, heterocyclyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(6a), or R⁷and R⁸ can be taken together with the nitrogen to which they areattached to form a monocyclic or bicyclic heterocycle with 3-7 membersin each ring and optionally containing, in addition to the nitrogen, oneor two additional heteroatoms selected from N, O and S, said monocyclicor bicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a);

R^(a) is (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, aryl, or heterocyclyl;

R^(b) is independently: H, (C₁-C₆)alkyl, aryl, heterocyclyl,(C₃-C₆)cycloalkyl, (C═O)_(a)O_(b)(C₁-C₆)alkyl, or S(O)_(m)R^(a); and

R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸,said alkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a second embodiment of this invention, the inhibitors of Akt activityare illustrated by the Formula A, wherein:

is selected from:

Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H;

and all other substituents and variables are as defined in the firstembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a third embodiment of this invention, the inhibitors of Akt activityare illustrated by the Formula B:

wherein:

Ring Y is cyclobutyl, said cyclobutyl is substituted with R^(x), andfurther optionally substituted with one or more substituents selectedfrom: (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, CO₂H, halo, CN, OHand NR⁷R⁸, said alkyl, cycloalkyl, and alkoxy is optionally substitutedwith one or more substituents selected from halo, CN, OH and NR⁷R⁸;

p is 0, 1 or 2;

R² is independently selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, CO₂H,halo, OH and NH₂;

and all other substituents and variables are as defined in the secondembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a fourth embodiment the inhibitors of the instant invention areillustrated by the Formula C:

wherein:

a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; p is 0, 1 or 2;

R² is independently selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, CO₂H,halo, OH and NH₂;

R¹ is selected from: H, oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)_(m)NR⁷R⁸, SH,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶;

R⁶ is: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo, CN, OH,O_(b)(C₁-C₆)perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO, (N═O)R⁷R⁸,S(O)_(m)NR⁷R⁸, SH, S(O)_(m)—(C₁-C₁₀)alkyl or(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, said alkyl, aryl, alkenyl, alkynyl,heterocyclyl, and cycloalkyl optionally substituted with one or moresubstituents selected from R^(6a);

R^(6a) is selected from: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,O_(a)(C₁-C₃)perfluoroalkyl, (C₀-C₆)alkylene-S(O)_(m)R^(a), SH, oxo, OH,halo, CN, (C₂-C₁₀)alkenyl, (C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl,(C₀-C₆)alkylene-aryl, (C₀-C₆)alkylene-heterocyclyl,(C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a), (C₀-C₆)alkylene-CO₂R^(a), C(O)H,and (Q-C₆)alkylene-CO₂H, said alkyl, alkenyl, alkynyl, cycloalkyl, aryl,and heterocyclyl is optionally substituted with up to three substituentsselected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, and N(R^(b))₂;

R⁷ and R⁸ are independently selected from: H,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)-heterocyclyl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, SH, SO₂R^(a), and (C═O)_(a)NR^(b) ₂, said alkyl,cycloalkyl, aryl, heterocyclyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(6a), or R⁷and R⁸ can be taken together with the nitrogen to which they areattached to form a monocyclic or bicyclic heterocycle with 3-7 membersin each ring and optionally containing, in addition to the nitrogen, oneor two additional heteroatoms selected from N, O and S, said monocyclicor bicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a);

R^(a) is (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, aryl, or heterocyclyl; and

R^(b) is independently: H, (C₁-C₆)alkyl, aryl, heterocyclyl,(C₃-C₆)cycloalkyl, (C═O)_(a)O_(b)(C₁-C₆)alkyl, or S(O)_(m)R^(a);

R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸,said alkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸;

R^(y) is selected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,(C₁-C₆)alkoxy, halo, CN and OH, said alkyl, cycloalkyl, and alkoxy isoptionally substituted with one or more substituents selected from halo,CN, OH and NR⁷R⁸; and

Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a fifth embodiment the inhibitors of the instant invention areillustrated by the Formula D:

wherein:

a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; p is 0, 1 or 2;

R² is independently selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, CO₂H,halo, OH and NH₂;

R¹ is selected from: H, oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)_(m)NR⁷R⁸, SH,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶;

R⁶ is: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo, CN, OH,O_(b)(C₁-C₆)perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO, (N═O)R⁷R⁸,S(O)_(m)NR⁷R⁸, SH, S(O)_(m)—(C₁-C₁₀)alkyl or(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, said alkyl, aryl, alkenyl, alkynyl,heterocyclyl, and cycloalkyl optionally substituted with one or moresubstituents selected from R^(6a);

R^(6a) is selected from: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,O_(a)(C₁-C₃)perfluoroalkyl, (C₀-C₆)alkylene-S(O)_(m)R^(a), SH, oxo, OH,halo, CN, (C₂-C₁₀)alkenyl, (C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl,(C₀-C₆)alkylene-aryl, (C₀-C₆)alkylene-heterocyclyl,(C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a), (C₀-C₆)alkylene-CO₂R^(a), C(O)H,and (C₀-C₆)alkylene-CO₂H, said alkyl, alkenyl, alkynyl, cycloalkyl,aryl, and heterocyclyl is optionally substituted with up to threesubstituents selected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, and N(R^(b))₂;

R⁷ and R⁸ are independently selected from: H,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)(C₃-C₉)cycloalkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)-heterocyclyl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, SH, SO₂R^(a), and (C═O)_(a)NR^(b) ₂, said alkyl,cycloalkyl, aryl, heterocyclyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(6a), or R⁷and R⁸ can be taken together with the nitrogen to which they areattached to form a monocyclic or bicyclic heterocycle with 3-7 membersin each ring and optionally containing, in addition to the nitrogen, oneor two additional heteroatoms selected from N, O and S, said monocyclicor bicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a);

R^(a) is (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, aryl, or heterocyclyl; and

R^(b) is independently: H, (C₁-C₆)alkyl, aryl, heterocyclyl,(C₃-C₆)cycloalkyl, (C═O)_(a)O_(b)(C₁-C₆)alkyl, or S(O)_(m)R^(a);

R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸,said alkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸;

R^(y) is selected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,(C₁-C₆)alkoxy, halo, CN and OH, said alkyl, cycloalkyl, and alkoxy isoptionally substituted with one or more substituents selected from halo,CN, OH and NR⁷R⁸; and

Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a sixth embodiment of this invention, the inhibitors of Akt activityare illustrated by the Formula E:

wherein:

a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; p is 0, 1 or 2;

R² is independently selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, CO₂H,halo, OH and NH₂;

R¹ is selected from: H, oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)_(m)NR⁷R⁸, SH,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶;

R⁶ is: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo, CN, OH,O_(b)(C₁-C₆)perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO, (N═O)R⁷R⁸,S(O)_(m)NR⁷R⁸, SH, S(O)_(m)—(C₁-C₁₀)alkyl or(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, said alkyl, aryl, alkenyl, alkynyl,heterocyclyl, and cycloalkyl optionally substituted with one or moresubstituents selected from R^(6a);

R^(6a) is selected from: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,O_(a)(C₁-C₃)perfluoroalkyl, (C₀-C₆)alkylene-S(O)_(m)R^(a), SH, oxo, OH,halo, CN, (C₂-C₁₀)alkenyl, (C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl,(C₀-C₆)alkylene-aryl, (C₀-C₆)alkylene-heterocyclyl,(C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a), (C₀-C₆)alkylene-CO₂R^(a), C(O)H,and (C₀-C₆)alkylene-CO₂H, said alkyl, alkenyl, alkynyl, cycloalkyl,aryl, and heterocyclyl is optionally substituted with up to threesubstituents selected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, and N(R^(b))₂;

R⁷ and R⁸ are independently selected from: H,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)-heterocyclyl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, SH, SO₂R^(a), and (C═O)_(a)NR^(b) ₂, said alkyl,cycloalkyl, aryl, heterocyclyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(6a), or R⁷and R⁸ can be taken together with the nitrogen to which they areattached to form a monocyclic or bicyclic heterocycle with 3-7 membersin each ring and optionally containing, in addition to the nitrogen, oneor two additional heteroatoms selected from N, O and S, said monocyclicor bicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a);

R^(a) is (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, aryl, or heterocyclyl; and

R^(b) is independently: H, (C₁-C₆)alkyl, aryl, heterocyclyl,(C₃-C₆)cycloalkyl, (C═O)_(a)O_(b)(C₁-C₆)alkyl, or S(O)_(m)R^(a);

R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸,said alkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸;

R^(y) is selected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,(C₁-C₆)alkoxy, halo, CN and OH, said alkyl, cycloalkyl, and alkoxy isoptionally substituted with one or more substituents selected from halo,CN, OH and NR⁷R⁸; and

Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a seventh embodiment of this invention, the inhibitors of Aktactivity are illustrated by the Formula C-1:

wherein:

R¹ is imidazolyl, triazolyl, or pyrimidyl, said imidazolyl, triazolyl,and pyrimidyl is optionally substituted with one or more (C₁-C₆)alkyl;

R⁷ and R⁸ are independently selected from: H, and (C₁-C₆)alkyl;

R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸,said alkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸;

R^(y) is selected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,(C₁-C₆)alkoxy, halo, CN and OH, said alkyl, cycloalkyl, and alkoxy isoptionally substituted with one or more substituents selected from halo,CN, OH and NR⁷R⁸; and

Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In an eighth embodiment of this invention, the inhibitors of Aktactivity are illustrated by the Formula D-1:

wherein:

R¹ is pyridyl, said pyridyl is optionally substituted with one or more(C₁-C₆)alkyl;

R⁷ and R⁸ are independently selected from: H, and (C₁-C₆)alkyl;

R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸,said alkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸;

R^(y) is selected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,(C₁-C₆)alkoxy, halo, CN and OH, said alkyl, cycloalkyl, and alkoxy isoptionally substituted with one or more substituents selected from halo,CN, OH and NR⁷R⁸; and

Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a ninth embodiment of this invention, the inhibitors of Akt activityare illustrated by the Formula D-1:

wherein:

R¹ is (C₁-C₆)alkyl, NR⁷R⁸, phenyl, pyridyl, pyrimidyl, pyrazinyl,azetidinyl, piperidinyl, or morpholinyl, said alkyl, phenyl, pyridyl,pyrimidyl, pyrazinyl, azetidinyl, piperidinyl, and morpholinyl isoptionally substituted with one or more substituents selected from:(C₁-C₆)alkyl, halo, OH, and NR⁷R⁸, said alkyl is optionally substitutedwith one or more substituents selected from: halo, and OH;

R⁷ and R⁸ are independently selected from: H, and (C₁-C₆)alkyl;

R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸,said alkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸;

R^(y) is selected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,(C₁-C₆)alkoxy, halo, CN and OH, said alkyl, cycloalkyl, and alkoxy isoptionally substituted with one or more substituents selected from halo,CN, OH and NR⁷R⁸; and

Bond:

is a single or double bond, provided that when 10 is oxo, then said bondis a single bond and adjacent N bears H;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a tenth embodiment of this invention, the inhibitors of Akt activityare illustrated by the Formula E-1:

wherein:

R¹ is pyrimidyl, said pyrimidyl is optionally substituted with one ormore (C₁-C₆)alkyl;

R⁷ and R⁸ are independently selected from: H, and (C₁-C₆)alkyl;

R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸,said alkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸;

R^(y) is selected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,(C₁-C₆)alkoxy, halo, CN and OH, said alkyl, cycloalkyl, and alkoxy isoptionally substituted with one or more substituents selected from halo,CN, OH and NR⁷R⁸; and

Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

Specific compounds of the instant invention include:

-   (1)    cis-3-amino-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (1-14);-   (2)    trans-3-amino-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (1-15);-   (3)    cis-3-amino-1-methyl-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (1-16);-   (4)    trans-3-amino-1-methyl-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (1-17);-   (5)    3,3-dimethoxy-1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (1-18);-   (6)    trans-3-amino-1-methyl-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-7);-   (7)    trans-3-fluoro-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (2-8);-   (8)    cis-3-fluoro-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (2-9);-   (9)    3,3-difluoro-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (2-10);-   (10)    trans-3-amino-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-11);-   (11)    cis-3-amino-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-12);-   (12)    trans-3-methoxy-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (2-13);-   (13)    trans-3-amino-1-cyclopropyl-3-{-4-[9-phenyl-3-(2-pyrimidinyl)-[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-14);-   (14)    cis-3-amino-1-cyclopropyl-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo    naphthyridin-8-yl]phenyl}cyclobutanol (2-15);-   (15) 3-fluoro-3-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)    [1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (2-16);-   (16) trans-3-amino-1-methyl-3-{4-[2-phenyl-9-(3    pyridinyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-7);-   (17)    2,2,2-trifluoro-N-(3-hydroxy-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide    (4-3);-   (18) N-(3-(dimethylamino)-1-{4-[9-phenyl-3-(2-pyrimidinyl)    [1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide    (4-4);-   (19)    N³,N³-dimethyl-1-{-4-[9-phenyl-3-(2-pyrimidinyl)-[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-1,3-cyclobutanediamine    (4-5);-   (20)    N³-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-1,3-cyclobutanediamine    (4-6);-   (21)    N-(3,3-difluoro-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide    (4-7);-   (22)    3-(methylamino)-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (4-9);-   (23)    N-(3-cyano-3-hydroxy-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide    (4-10);-   (24)    3-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (4-13);-   (25)    3-amino1-(hydroxymethyl)-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo    naphthyridin-8-yl]phenyl}cyclobutanol (445);-   (26)    3,3-difluoro-1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl])phenyl}cyclobutanamine    (5-12);-   (27) 3,3-difluoro-1-{4-[9-phenyl-3-(trifluoromethyl)    [1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (5-13);-   (28)    3,3-difluoro-1-{-4-[9-phenyl-3-(1H-1,2,3-triazol-4-yl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (5-14);-   (29)    3,3-difluoro-1-{4-[3-(1H-indazol-3-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (5-15);-   (30)    3,3-difluoro-1-{4-[3-(5-methyl-1H-1,2,4-triazol-3-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (5-16);-   (31)    8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridine-3-carboxamide    (5-17);-   (32)    8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-amine    (5-19);-   (33)    8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-ol    (5-21);-   (34)    3-amino-3-{4-[9-phenyl-2-(2-pyrimidinyl)[1,2,4]triazolo[5,1-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (6-1);-   (35) trans-3-amino-1-ethyl-3-{4-[9-phenyl-3-(pyrimidin-2-yl)    [1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-17);-   (36)    trans-3-amino-1-methyl-3-{4-[3-(pyrimidin-2-yl)-9-(thiophen-2-yl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-18);-   (37) trans-3-amino-1-methyl-3-{4-[9-phenyl-3-(pyrimidin-2-yl)    [1,2,4]triazolo[4′,3′:1,2]pyrido[3,4-b]pyrazin-8-yl]phenyl}cyclobutanol    (2-19);-   (38)    8-[4-(trans-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-ol    (2-21);-   (39)    trans-3-amino-1-cyclopropyl-3-{4-[9-(3,4-difluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-8);-   (40)    trans-3-amino-1-cyclopropyl-3-[4-(2,9-diphenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]cyclobutanol    (3-9);-   (41)    trans-3-amino-3-{4-[9-(4-chlorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol    (3-10);-   (42)    3-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol    (3-11);-   (43)    trans-3-amino-1-cyclopropyl-3-{4-[9-(4-methylphenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-12);-   (44)    trans-3-amino-1-methyl-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-13);-   (45)    4-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol    (3-14);-   (46)    trans-3-amino-1-cyclopropyl-3-{4-[9-(4-fluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-15);-   (47)    trans-3-amino-1-cyclopropyl-3-{4-[9-(3-fluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-16);-   (48)    3-amino-3-{4-[9-(2-aminopyrimidin-5-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol    (3-17);-   (49)    3-{3-[4-(trans-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol    (3-18);-   (50)    trans-3-amino-3-{4-[9-(6-aminopyridin-3-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol    (3-19);-   (51)    trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-20);-   (52)    trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(pyridin-3-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-21);-   (53)    trans-3-amino-1-ethyl-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-22);-   (54)    trans-3-amino-1-cyclopropyl-3-{4-[9-(morpholin-4-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-23);-   (55)    trans-3-amino-3-[4-(9-ethyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-methylcyclobutanol    (3-24);-   (56)    trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(pyrimidin-5-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-25);-   (57)    trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(piperidin-1-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-27);-   (58)    trans-3-amino-3-{4-[9-(2-aminopyridin-4-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-methylcyclobutanol    (3-28);-   (59)    trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(pyrazin-2-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-29);-   (60)    (3-amino-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutyl)methanol    (3-30);-   (61)    trans-3-amino-1-cyclopropyl-3-{4-[9-(difluoromethyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-31);-   (62)    trans-3-amino-3-{4-[9-(difluoromethyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-methylcyclobutanol    (3-32);-   (63)    trans-3-amino-3-{4-[9-(3-chlorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol    (3-33);-   (64)    trans-3-amino-1-cyclopropyl-3-{4-[9-(2-fluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-34);-   (65) cis-3-amino-1-methyl-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)    [1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-35);-   (66)    trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(trifluoromethyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-36);-   (67)    trans-3-amino-1-cyclopropyl-3-{4-[9-(3-methylphenyl)-2-phenyl][1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol    (3-37);-   (68)    cis-3-amino-1-methyl-3-{4-[9-(morpholin-4-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-38);-   (69)    (3-amino-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutyl)methanol    (3-39);-   (70)    3-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl}benzonitrile    (3-40);-   (71)    cis-3-amino-1-methyl-3-{4-[2-phenyl-9-(pyrimidin-5-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-41);-   (72)    2-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol    (3-42);-   (73)    trans-3-amino-3-[4-(9-tert-butyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropyl    cyclobutanol (3-44);-   (74)    trans-3-amino-1-cyclopropyl-3-(4-{2-phenyl-9-[3-(trifluoromethyl)phenyl][1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol    (3-45);-   (75)    trans-3-amino-1-cyclopropyl-3-(4-{9-[4-(methylsulfonyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol    (3-46);-   (76)    trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(1H-pyrrolo[2,3-b]pyridin-4-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-47);-   (77) 3,3-difluoro-1-{4-[2-phenyl-9-(1H-1,2,4-triazol-3-yl)    [1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanamine    (3-48);-   (78)    trans-3-amino-1-cyclopropyl-3-{4-[9-(2-hydroxypropan-2-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-49);-   (79)    trans-3-amino-1-cyclopropyl-3-[4-(9-methyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]cyclobutanol    (3-50);-   (80)    trans-3-amino-1-cyclopropyl-3-{4-[9-(dimethylamino)-2-phenyl-[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (8-5);-   (81)    trans-3-amino-1-cyclopropyl-3-{4-[9-(methylamino)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (8-6);-   (82)    trans-3-amino-3-[4-(9-amino-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol    (8-7);-   (83)    trans-3-amino-3-{4-[9-(azetidin-1-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol    (8-8);-   (84)    trans-3-amino-3-[4-(9-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol    (8-9);-   (85)    trans-3-amino-1-cyclopropyl-3-{4-[9-(methylsulfanyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (8-10);-   (86)    trans-3-amino-1-cyclopropyl-3-[4-(9-methoxy-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]cyclobutanol    (8-11);-   (87)    3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-ol    (8-12);-   (88)    3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-carboxamide    (9-4);-   (89)    trans-3-amino-1-cyclopropyl-3-(4-{9-[(dimethylamino)methyl]-2-phenyl-[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol    (10-5);-   (90)    N-({3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl}methyl)acetamide    (10-6); and-   (91)    3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-N-ethyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-carboxamide    (11-5);    or a pharmaceutically acceptable salt or stereoisomer thereof.

The compounds of the present invention may have asymmetric centers,chiral axes, and chiral planes (as described in: E. L. Eliel and S. H.Wilen, Stereochemistry of Carbon Compounds, John Wiley & Sons, New York,1994, pages 1119-1190), and occur as racemates, racemic mixtures, and asindividual diastereomers, with all possible isomers and mixturesthereof, including optical isomers, all such stereoisomers beingincluded in the present invention.

In addition, the compounds disclosed herein may exist as tautomers andboth tautomeric forms are intended to be encompassed by the scope of theinvention, even though only one tautomeric structure is depicted. Forexample the following is within the scope of the instant invention:

Tetrazoles exist as a mixture of 1H/2H tautomers. The tautomeric formsof the tetrazol moiety are also within the scope of the instantinvention.

This invention is also intended to encompass pro-drugs of the compoundsdisclosed herein. A prodrug of any of the compounds can be made usingwell known pharmacological techniques.

When any variable (e.g. R², etc.) occurs more than one time in anyconstituent, its definition on each occurrence is independent at everyother occurrence. Also, combinations of substituents and variables arepermissible only if such combinations result in stable compounds. Linesdrawn into the ring systems from substituents represent that theindicated bond may be attached to any of the substitutable ring atoms.If the ring system is bicyclic, it is intended that the bond be attachedto any of the suitable atoms on either ring of the bicyclic moiety.

It is understood that one or more silicon (Si) atoms can be incorporatedinto the compounds of the instant invention in place of one or morecarbon atoms by one of ordinary skill in the art to provide compoundsthat are chemically stable and that can be readily synthesized bytechniques known in the art from readily available starting materials.Carbon and silicon differ in their covalent radius leading todifferences in bond distance and the steric arrangement when comparinganalogous C-element and Si-element bonds. These differences lead tosubtle changes in the size and shape of silicon-containing compoundswhen compared to carbon. One of ordinary skill in the art wouldunderstand that size and shape differences can lead to subtle ordramatic changes in potency, solubility, lack of off target activity,packaging properties, and so on. (Diass, J. O. et al. Organometallics(2006) 5:1188-1198; Showell, G. A. et al. Bioorganic & MedicinalChemistry Letters (2006) 16:2555-2558).

It is understood that substituents and substitution patterns on thecompounds of the instant invention can be selected by one of ordinaryskill in the art to provide compounds that are chemically stable andthat can be readily synthesized by techniques known in the art, as wellas those methods set forth below, from readily available startingmaterials. If a substituent is itself substituted with more than onegroup, it is understood that these multiple groups may be on the samecarbon or on different carbons, so long as a stable structure results.The phrase “optionally substituted with one or more substituents” shouldbe taken to be equivalent to the phrase “optionally substituted with atleast one substituent” and in such cases the preferred embodiment willhave from zero to four substituents, and the more preferred embodimentwill have from zero to three substituents.

As used herein, “alkyl” is intended to include both branched andstraight-chain saturated aliphatic hydrocarbon groups having thespecified number of carbon atoms. For example, C₁-C₁₀, as in“(C₁-C₁₀)alkyl” is defined to include groups having 1, 2, 3, 4, 5, 6, 7,8, 9 or 10 carbons in a linear or branched arrangement. For example,“(C₁-C₁₀)alkyl” specifically includes methyl, ethyl, n-propyl, i-propyl,n-butyl, t-butyl, i-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl,and so on.

The term “cycloalkyl” means a monocyclic saturated aliphatic hydrocarbongroup having the specified number of carbon atoms. For example,“cycloalkyl” includes cyclopropyl, methyl-cyclopropyl,2,2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl, cyclohexyl, and so on.

“Alkoxy” represents either a cyclic or non-cyclic alkyl group ofindicated number of carbon atoms attached through an oxygen bridge.“Alkoxy” therefore encompasses the definitions of alkyl and cycloalkylabove.

If no number of carbon atoms is specified, the term “alkenyl” refers toa non-aromatic hydrocarbon radical, straight, branched or cyclic,containing from 2 to 10 carbon atoms and at least one carbon to carbondouble bond. Preferably one carbon to carbon double bond is present, andup to four non-aromatic carbon-carbon double bonds may be present. Thus,“(C₂-C₁₀)alkenyl” means an alkenyl radical having from 2 to 10 carbonatoms. Alkenyl groups include ethenyl, propenyl, butenyl,2-methylbutenyl and cyclohexenyl. The straight, branched or cyclicportion of the alkenyl group may contain double bonds and may besubstituted if a substituted alkenyl group is indicated.

The term “alkynyl” refers to a hydrocarbon radical straight, branched orcyclic, containing from 2 to 10 carbon atoms and at least one carbon tocarbon triple bond. Up to three carbon-carbon triple bonds may bepresent. Thus, “(C₂-C₁₀)alkynyl” means an alkynyl radical having from 2to 10 carbon atoms. Alkynyl groups include ethynyl, propynyl, butynyl,3-methylbutynyl and so on. The straight, branched or cyclic portion ofthe alkynyl group may contain triple bonds and may be substituted if asubstituted alkynyl group is indicated.

In certain instances, substituents may be defined with a range ofcarbons that includes zero, such as (C₀-C₆)alkylene-aryl. If aryl istaken to be phenyl, this definition would include phenyl itself as wellas —CH₂Ph, —CH₂CH₂Ph, CH(CH₃)CH₂CH(CH₃)Ph, and so on.

As used herein, “aryl” is intended to mean any stable monocyclic orbicyclic carbon ring of up to 7 atoms in each ring, wherein at least onering is aromatic. Examples of such aryl elements include phenyl,naphthyl, tetrahydro-naphthyl, indanyl and biphenyl. In cases where thearyl substituent is bicyclic and one ring is non-aromatic, it isunderstood that attachment is via the aromatic ring.

The term heteroaryl, as used herein, represents a stable monocyclic orbicyclic ring of up to 7 atoms in each ring, wherein at least one ringis aromatic and contains from 1 to 4 heteroatoms selected from the groupconsisting of O, N and S. Heteroaryl groups within the scope of thisdefinition include but are not limited to: acridinyl, carbazolyl,cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, furanyl,thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl,oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl,pyrimidinyl, pyrrolyl, tetrahydroquinoline. As with the definition ofheterocycle below, “heteroaryl” is also understood to include theN-oxide derivative of any nitrogen-containing heteroaryl. In cases wherethe heteroaryl substituent is bicyclic and one ring is non-aromatic orcontains no heteroatoms, it is understood that attachment is via thearomatic ring or via the heteroatom containing ring, respectively. Suchheteroaryl moieties for substituent Q include but are not limited to:2-benzimidazolyl, 2-quinolinyl, 3-quinolinyl, 4-quinolinyl,1-isoquinolinyl, 3-isoquinolinyl and 4-isoquinolinyl.

The term “heterocycle” or “heterocyclyl” as used herein is intended tomean a 3- to 10-membered aromatic or non-aromatic heterocycle containingfrom 1 to 4 heteroatoms selected from the group consisting of O, N andS, and includes bicyclic groups. “Heterocyclyl” therefore includes theabove mentioned heteroaryls, as well as dihydro and tetrahydro analogsthereof. Further examples of “heterocyclyl” include, but are not limitedto the following: benzoimidazolyl, benzoimidazolonyl, benzofuranyl,benzofurazanyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl,benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl, furanyl, imidazolyl,indolinyl, indolyl, indolazinyl, indazolyl, isobenzofuranyl, isoindolyl,isoquinolyl, isothiazolyl, isoxazolyl, naphthpyridinyl, oxadiazolyl,oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl,pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl,pyrimidyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl,tetrahydropyranyl, tetrazolyl, tetrazolopyridyl, thiadiazolyl,thiazolyl, thienyl, triazolyl, azetidinyl, 1,4-dioxanyl,hexahydroazepinyl, piperazinyl, piperidinyl, pyridin-2-onyl,pyrrolidinyl, morpholinyl, thiomorpholinyl, dihydrobenzimidazolyl,dihydrobenzofuranyl, dihydrobenzothiophenyl, dihydrobenzoxazolyl,dihydrofuranyl, dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl,dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl,dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl,dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl, andN-oxides thereof. Attachment of a heterocyclyl substituent can occur viaa carbon atom or via a heteroatom.

As appreciated by those of skill in the art, “halo” or “halogen” as usedherein is intended to include chloro (Cl), fluoro (F), bromo (Br) andiodo (I).

In an embodiment of Formula A and B, the moiety illustrated by theformula:

includes the following structures:

In another embodiment of Formula A and B, the moiety illustrated by theformula:

includes the following structures:

In another embodiment of Formula A and B, the moiety illustrated by theformula:

In yet another embodiment of Formula A and 13, the moiety illustrated bythe formula:

In an embodiment of Formula A and B, Ring Z is selected from: phenyl andheterocyclyl.

In another embodiment, Ring Z is selected from:

In yet another embodiment, Ring Z is phenyl.

In an embodiment, p is independently 0, 1, 2 or 3.

In a further embodiment, p is independently 0, 1 or 2.

In another embodiment, p is independently 1.

In yet another embodiment, p is independently 0.

In an embodiment of Formula A, B, C, D and E, R² is selected from: H andhalogen.

In another embodiment of Formula A, B, C, D and E, R² is selected from:H and F.

In an embodiment of Formula A, B, C, D and E, R¹ is selected from: oxo,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)-aryl,(C═O)_(a)O_(b)(C₂-C₁₀)alkenyl, (C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H,halo, OH, O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)₂NR⁷R⁸, SH, S(O)₂—(C₁-C₁₀)alkyl and(C═O)_(a)O_(b)-heterocyclyl, said alkyl, aryl, alkenyl, alkynyl,cycloalkyl, and heterocyclyl is optionally substituted with R⁶; R⁶ is:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C═O)_(a)O_(b)heterocyclyl, halo, OH, O_(b)(C₁-C₆)perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸,oxo, or (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl; R⁷ and R⁸ in the group of(C═O)_(a)NR⁷R⁸, S(O)₂NR⁷R⁸, or O_(a)(C═O)_(b)NR⁷R⁸, are independentlyselected from: H, and (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, a is 0 or 1; and b is0 or 1.

In another embodiment of Formula A, B, C, D and E, R¹ is selected from:oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)-aryl, (C═O)_(a)NR⁷R⁸,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, and (C═O)_(a)O_(b)-heterocyclyl, saidalkyl, aryl, cycloalkyl, and heterocyclyl is optionally substituted withone or more substituents selected from R⁶; R⁶ is:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C═O)_(a)O_(b)heterocyclyl, halo, OH, O_(b)(C₁-C₆)perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸,oxo, or (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl; R⁷ and R⁸ in the group of(C═O)_(a)NR⁷R⁸, or O_(a)(C═O)_(b)NR⁷R⁸, are independently selected from:H, and (C═O)_(a)O_(b)(C₁-C₁₀)alkyl; a is 0 or 1; and b is 0 or 1.

In another embodiment of Formula A, B, C, D and E, R¹ is (C₁-C₆)alkyl,OH, (C═O)_(a)NR⁷R⁸, phenyl, imidazolyl, pyrazolyl, triazolyl, pyridyl,pyrimidyl, pyrazinyl, pyridazinyl, azetidinyl, piperidinyl, ormorpholinyl, said alkyl, phenyl, imidazolyl, pyrazolyl, triazolyl,pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, azetidinyl, piperidinyl, andmorpholinyl is optionally substituted with one or more substituentsselected from: (C₁-C₆)alkyl, halo, OH, and (C═O)_(b)NR⁷R⁸, said alkyl isoptionally substituted with one or more substituents selected from:halo, and OH; R⁷ and R⁸ in the group of (C═O)_(a)NR⁷R⁸, or(C═O)_(b)NR⁷R⁸, are independently selected from: H, and (C₁-C₆)alkyl; ais 0 or 1; and b is 0 or 1.

In another embodiment of Formula A, B, C, D and E, R¹ is imidazolyl,pyridyl, or pyrimidyl, said imidazolyl, triazolyl, pyridyl, andpyrimidyl is optionally substituted with one or more (C₁-C₆)alkyl.

In another embodiment of Formula A, B, C, D and E, R¹ is (C₁-C₆)alkyl,NR⁷R⁸, phenyl, pyridyl, pyrimidyl, pyrazinyl, azetidinyl, piperidinyl,or morpholinyl, said alkyl, phenyl, pyridyl, pyrimidyl, pyrazinyl,azetidinyl, piperidinyl, and morpholinyl is optionally substituted withone or more substituents selected from: (C₁-C₆)alkyl, halo, OH, andNR⁷R⁸, said alkyl is optionally substituted with one or moresubstituents selected from: halo, and OH; R⁷ and R⁸ are independentlyselected from: H, and (C₁-C₆)alkyl.

In an embodiment of Formula A, B, C, D and E, (C₁-C₆)alkyl for R¹ ismethyl or ethyl, which is optionally substituted with OH.

In another embodiment of Formula A, B, C, D, D-1 and E, (C₁-C₆)alkyl forR¹ is methyl, ethyl, or propyl, which is optionally substituted with oneor more halo, preferably F.

In an embodiment of Formula A, B, C, D and E, (C═O)_(a)NR⁷R⁸ for 10 is(C═O)NH₂.

In another embodiment of Formula A, B, C, D and E, (C═O)_(a)NR⁷R⁸ for R¹is NH₂, or NHCH₃.

In an embodiment of Formula D-1, NR⁷R⁸ for R¹ is NH₂, or NHCH₃.

In an embodiment of Formula A, B, C, D, E, C-1, D-1, and E-1,(C₁-C₆)alkyl as the substituents for R¹ is methyl or ethyl, preferablymethyl.

In an embodiment of Formula A, B, C, D and E, halo as the substituentsfor R¹ is F.

In an embodiment of Formula A, B, C, D and E, R⁷ and R⁸ in the formula:

are independently selected from: H, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, (C═O)_(a)O_(b)-heterocyclyl, SO₂R^(a),and (C═O)_(a)NR^(b) ₂, said alkyl, cycloalkyl, aryl, and heterocyclyl isoptionally substituted with one or more substituents selected fromR^(6a); R^(6a) is selected from: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, OH, halo,CN, (C₀-C₆)alkylene-aryl, and (C₀-C₆)alkylene-heterocyclyl, said alkyl,aryl, and heterocyclyl is optionally substituted with up to threesubstituents selected from R^(b), halogen, and oxo; R^(a) is(C₁-C₆)alkyl, aryl, or heterocyclyl, said alkyl, aryl, and heterocyclylis optionally substituted with one or more substituents selected fromR^(c); R^(b) is independently: H, (C₁-C₆)alkyl, aryl, or heterocyclyl,said alkyl, aryl, and heterocyclyl is optionally substituted with one ormore aryl; R^(c) is independently: aryl, halo, or CN, said aryl isoptionally substituted with one or more CN; a is 0 or 1; and b is 0 or1.

In another embodiment of Formula A, B, C, D and E, both R⁷ and R⁸ in theformula:

are H.

In an embodiment of Formula A, Ring Y is cyclobutyl, said cyclobutyl issubstituted with R^(x), and further optionally substituted with one ormore substituents selected from: (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,(C₁-C₆)alkoxy, CO₂H, halo, CN, OH and NR⁷R⁸, said alkyl, cycloalkyl, andalkoxy is optionally substituted with one or more substituents selectedfrom halo, CN, OH and NR⁷R⁸; R^(x) is selected from: (C₁-C₆)alkyl,(C₁-C₆)alkoxy, halo, OH and NR⁷R⁸, said alkyl, and alkoxy is optionallysubstituted with one or more substituents selected from halo, CN, OH andNR⁷R⁸; and R⁷ and R⁸ in the group of NR⁷R⁸ as R^(x) or the substituents,are independently selected from: H, and (C₁-C₆)alkyl.

In another embodiment of Formula A and B, Ring Y is cyclobutyl, saidcyclobutyl is substituted with R^(x), and further optionally substitutedwith one or more substituents selected from: (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, halo, CN and OH, said alkyl,cycloalkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸; R^(x) is selectedfrom: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸, said alkyl, andalkoxy is optionally substituted with one or more substituents selectedfrom halo, CN, OH and NR⁷R⁸; and R⁷ and R⁸ in the group of NR⁷R⁸ asR^(x) or the substituents, are independently selected from: H, and(C₁-C₆)alkyl.

In another embodiment of Formula A and B, Ring Y is cyclobutyl, saidcyclobutyl is substituted with R^(x); and further optionally substitutedwith one or more substituents selected from: (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, and halo, said alkyl, and cycloalkyl is optionallysubstituted with one or more substituents selected from halo, CN, OH andNR⁷R⁸; R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, or OH,said alkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸; and R⁷ and R⁸ in thegroup of NR⁷R⁸ as the substituents, are independently selected from: H,and (C₁-C₆)alkyl.

In another embodiment of Formula A and B, Ring Y is cyclobutyl, saidcyclobutyl is substituted with R^(x); and further optionally substitutedwith one or more substituents selected from: (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, and halo, said alkyl, and cycloalkyl is optionallysubstituted with one or more substituents selected from halo, and OH;R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, or OH, saidalkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, and OH.

In another embodiment of Formula A and B, Ring Y is cyclobutyl, saidcyclobutyl is substituted with OH, and further optionally substitutedwith one substituent selected from: (C₁-C₆)alkyl, and (C₃-C₆)cycloalkyl,said alkyl, and cycloalkyl is optionally substituted with OH.

In yet another embodiment of Formula A and B, Ring Y is cyclobutyl, saidcyclobutyl is substituted with OH, and further optionally substitutedwith one substituent selected from: methyl and cyclopropyl, said methylis optionally substituted with OH.

In another embodiment of Formula A and B, Ring Y is cyclobutyl, saidcyclobutyl is substituted with halo, preferably F, and furtheroptionally substituted with one substituent selected from: (C₁-C₆)alkyl,and halo, said alkyl is optionally substituted with one or moresubstituents selected from halo, and OH.

In yet another embodiment of Formula A and B, Ring Y is cyclobutyl, saidcyclobutyl is substituted with halo, preferably F, and furtheroptionally substituted with one substituent selected from: methyl, andF.

In another embodiment of Formula A and B, Ring Y is cyclobutyl, saidcyclobutyl is substituted with (C₁-C₆)alkyl, preferably methyl.

In another embodiment of Formula A and B, Ring Y is cyclobutyl, saidcyclobutyl is substituted with (C₁-C₆)alkoxy, preferably methoxy.

In an embodiment of Formula C, D, E, C-1, D-1, and E-1, R^(x) isselected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, and OH, said alkyl,and alkoxy is optionally substituted with one or more substituentsselected from halo, CN, OH and NR⁷R⁸; R^(y) is selected from: H,(C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, and halo, said alkyl, cycloalkyl, andalkoxy is optionally substituted with one or more substituents selectedfrom halo, CN, OH and NR⁷R⁸; and R⁷ and R⁸ in the group of NR⁷R⁸ as thesubstituents, are independently selected from: H, and (C₁-C₆)alkyl.

In another embodiment of Formula C, D, E, C-1, D-1, and E-1, R^(x) isselected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, and OH, said alkyl,and alkoxy is optionally substituted with one or more substituentsselected from halo, and OH; R^(y) is selected from: H, (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, and halo, said alkyl, cycloalkyl, and alkoxy isoptionally substituted with one or more substituents selected from halo,and OH.

In another embodiment of Formula A, B, C, D, E, C-1, D-1, and E-1,(C₁-C₆)alkyl for R^(x) is methyl, or ethyl, preferably methyl.

In another embodiment of Formula A, B, C, D, E, C-1, D-1, and E-1,(C₁-C₆)alkoxy for R^(x) is methoxy or ethoxy, preferably methoxy.

In another embodiment of Formula A, B, C, D, E, C-1, D-1, and E-1, halofor R^(x) is F.

In another embodiment of Formula C, D, E, C-1, D-1, and E-1, R^(x) isOH; and R^(y) is selected from: H, (C₁-C₆)alkyl, and (C₃-C₆)cycloalkyl,said alkyl and cycloalkyl are optionally substituted with OH.

In yet another embodiment of Formula C, D, E, C-1, D-1, and E-1, R^(x)is OH; and R^(y) is selected from: H, methyl, and cyclopropyl, saidmethyl is optionally substituted with OH.

In another embodiment of Formula C, D, E, C-1, D-1, and E-1, R^(x) ishalo, preferably F; and R^(y) is selected from: H, (C₁-C₆)alkyl, andhalo, said alkyl is optionally substituted with one or more substituentsselected from halo, and OH.

In yet another embodiment of Formula C, D, E, C-1, D-1, and E-1, R^(x)is halo, preferably F; and R^(y) is selected from: H, methyl, and F.

In another embodiment of Formula C, D, E, C-1, D-1, and E-1, R^(x) is(C₁-C₆)alkyl, preferably methyl; and R^(y) is H.

In another embodiment of Formula C, D, E, C-1, D-1, and E-1, R^(x) is(C₁-C₆)alkoxy, preferably methoxy; and R^(y) is H.

Included in the instant invention is the free form of compounds ofFormula A, as well as the pharmaceutically acceptable salts andstereoisomers thereof. Some of the isolated specific compoundsexemplified herein are the protonated salts of amine compounds. The term“free form” refers to the amine compounds in non-salt form. Theencompassed pharmaceutically acceptable salts not only include theisolated salts exemplified for the specific compounds described herein,but also all the typical pharmaceutically acceptable salts of the freeform of compounds of Formula A. The free farm of the specific saltcompounds described may be isolated using techniques known in the art.For example, the free form may be regenerated by treating the salt witha suitable dilute aqueous base solution such as dilute aqueous NaOH,potassium carbonate, ammonia and sodium bicarbonate. The free forms maydiffer from their respective salt forms somewhat in certain physicalproperties, such as solubility in polar solvents, but the acid and basesalts are otherwise pharmaceutically equivalent to their respective freeforms for purposes of the invention.

The pharmaceutically acceptable salts of the instant compounds can besynthesized from the compounds of this invention which contain a basicor acidic moiety by conventional chemical methods. Generally, the saltsof the basic compounds are prepared either by ion exchangechromatography or by reacting the free base with stoichiometric amountsor with an excess of the desired salt-forming inorganic or organic acidin a suitable solvent or various combinations of solvents. Similarly,the salts of the acidic compounds are formed by reactions with theappropriate inorganic or organic base.

Thus, pharmaceutically acceptable salts of the compounds of thisinvention include the conventional non-toxic salts of the compounds ofthis invention as formed by reacting a basic instant compound with aninorganic or organic acid. For example, conventional non-toxic saltsinclude those derived from inorganic acids such as hydrochloric,hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like, aswell as salts prepared from organic acids such as acetic, propionic,succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic,pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic,salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic,methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic(TFA) and the like.

When the compound of the present invention is acidic, suitable“pharmaceutically acceptable salts” refers to salts prepared formpharmaceutically acceptable non-toxic bases including inorganic basesand organic bases. Salts derived from inorganic bases include aluminum,ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganicsalts, manganous, potassium, sodium, zinc and the like. Particularlypreferred are the ammonium, calcium, magnesium, potassium and sodiumsalts. Salts derived from pharmaceutically acceptable organic non-toxicbases include salts of primary, secondary and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion exchange resins, such as arginine, betainecaffeine, choline, N,N¹-dibenzylethylenediamine, diethylamin,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylaminetripropylamine, tromethamine and the like.

The preparation of the pharmaceutically acceptable salts described aboveand other typical pharmaceutically acceptable salts is more fullydescribed by Berg et al., “Pharmaceutical Salts,” J. Pharm. Sci.,1977:66:1-19.

It will also be noted that the compounds of the present invention arepotentially internal salts or zwitterions, since under physiologicalconditions a deprotonated acidic moiety in the compound, such as acarboxyl group, may be anionic, and this electronic charge might then bebalanced off internally against the cationic charge of a protonated oralkylated basic moiety, such as a quaternary nitrogen atom.

UTILITY

The compounds of the instant invention are inhibitors of the activity ofAkt and are thus useful in the treatment of cancer, in particularcancers associated with irregularities in the activity of Akt anddownstream cellular targets of Akt. Such cancers include, but are notlimited to, ovarian, pancreatic, breast and prostate cancer, as well ascancers (including glioblastoma) where the tumor suppressor PTEN ismutated (Cheng et al., Proc. Natl. Acad. Sci. (1992) 89:9267-9271; Chenget al., Proc. Natl. Acad. Sci. (1996) 93:3636-3641; Bellacosa et al.,Int. J. Cancer (1995) 64:280-285; Nakatani et al., J. Biol. Chem. (1999)274:21528-21532; Graff, Expert. Opin. Ther. Targets (2002) 6(1):103-113;and Yamada and Araki, J. Cell Science. (2001) 114:2375-2382; Mischel andCloughesy, Brain Pathol. (2003) 13(1):52-61). Cancers where Akt itselfis activated by gene amplification or mutations may also be treated bythe compounds. Human breast, colorectal and ovarian cancers where asomatic mutation in a pleckstrin homology domain (PH) of AKT1 (E17Kmutant; the glutamic acid (E) at position 17 of the amino acid sequenceof the PH domain of AKT1 is replaced by a lysine (K)) is reported(Carpten et al, Nature 448: 439-444 (2007)).

The compounds, compositions and methods provided herein are particularlydeemed useful for the treatment of cancer. Cancers that may be treatedby the compounds, compositions and methods of the invention include, butare not limited to: Cardiac: sarcoma (angiosarcoma, fibrosarcoma,rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma andteratoma; Lung: non-small cell lung, bronchogenic carcinoma (squamouscell, undifferentiated small cell, undifferentiated large cell,adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma,sarcoma, lymphoma, chondromatous hamartoma, mesothelioma;Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma,leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma,leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma,glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel(adenocarcinoma, lymphoma, carcinoid tumors, Karposi's sarcoma,leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel(adenocarcinoma, tubular adenoma, villous adenoma, hamartoma,leiomyoma), colon, colorectal, rectal; Genitourinary tract: kidney(adenocarcinoma, Wilm's tumor [nephroblastoma], lymphoma, leukemia),bladder and urethra (squamous cell carcinoma, transitional cellcarcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis(seminoma, teratoma, embryonal carcinoma, teratocarcinoma,choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma,fibroadenoma, adenomatoid tumors, lipoma); Liver: hepatoma(hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma,angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenicsarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma,chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cellsarcoma), multiple myeloma, malignant giant cell tumor chordoma,osteochronfroma (osteocartilaginous exostoses), benign chondroma,chondroblastoma, chondromyxofibroma, osteoid osteoma and giant celltumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma,osteitis deformans), meninges (meningioma, meningiosarcoma,gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma,germinoma [pinealoma], glioblastoma multiform, oligodendroglioma,schwannoma, retinoblastoma, congenital tumors), spinal cordneurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus(endometrial carcinoma), cervix (cervical carcinoma, pre-tumor cervicaldysplasia), ovaries (ovarian carcinoma [serous cystadenocarcinoma,mucinous cystadenocarcinoma, unclassified carcinoma], granulosa-thecalcell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignantteratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma,adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma,squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma),fallopian tubes (carcinoma); Hematologic: blood (myeloid leukemia [acuteand chronic], acute lymphoblastic leukemia, chronic lymphocyticleukemia, myeloproliferative diseases, multiple myeloma, myelodysplasticsyndrome), Hodgkin's disease, non-Hodgkin's lymphoma [malignantlymphoma]; Skin: malignant melanoma, basal cell carcinoma, squamous cellcarcinoma, Karposi's sarcoma, moles dysplastic nevi, lipoma, angioma,dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma.Thus, the term “cancerous cell” as provided herein, includes a cellafflicted by any one of the above-identified conditions.

Cancers that may be treated by the compounds, compositions and methodsof the invention include, but are not limited to: breast, prostate,colon, colorectal, lung, non-small cell lung, brain, testicular,stomach, pancreas, skin, small intestine, large intestine, throat, headand neck, oral, bone, liver, bladder, kidney, thyroid and blood.

Cancers that may be treated by the compounds, compositions and methodsof the invention include: breast, prostate, colon, ovarian, colorectal,lung and non-small cell lung.

Cancers that may be treated by the compounds, compositions and methodsof the invention include: breast, colon, (colorectal) and lung(non-small cell lung).

Cancers that may be treated by the compounds, compositions and methodsof the invention include: lymphoma and leukemia.

The compounds of the instant invention are useful for the treatment ofbreast cancer.

The compounds of the instant invention are useful for the treatment ofprostate cancer.

Akt signaling regulates multiple critical steps in angiogenesis.Shiojima and Walsh, Circ. Res. (2002) 90:1243-1250. The utility ofangiogenesis inhibitors in the treatment of cancer is known in theliterature, see J. Rak et al. Cancer Research, 55:4575-4580, 1995 andDredge et al., Expert Opin. Biol. Ther. (2002) 2(8):953-966, forexample. The role of angiogenesis in cancer has been shown in numeroustypes of cancer and tissues: breast carcinoma (G. Gasparini and A. L.Harris, J. Clin. Oncol., 1995, 13:765-782; M. Toi et al., Japan. J.Cancer Res., 1994, 85:1045-1049); bladder carcinomas (A. J. Dickinson etal., Br. J. Urol., 1994, 74:762-766); colon carcinomas (L. M. Ellis etal., Surgery, 1996, 120(5):871-878); and oral cavity tumors (J. K.Williams et al., Am. J. Surg., 1994, 168:373-380). Other cancersinclude, advanced tumors, hairy cell leukemia, melanoma, advanced headand neck, metastatic renal cell, non-Hodgkin's lymphoma, metastaticbreast, breast adenocarcinoma, advanced melanoma, pancreatic, gastric,glioblastoma, lung, ovarian, non-small cell lung, prostate, small celllung, renal cell carcinoma, various solid tumors, multiple myeloma,metastatic prostate, malignant glioma, renal cancer, lymphoma,refractory metastatic disease, refractory multiple myeloma, cervicalcancer, Kaposi's sarcoma, recurrent anaplastic glioma, and metastaticcolon cancer (Dredge et al., Expert Opin. Biol. Ther. (2002)2(8):953-966). Thus, the Akt inhibitors disclosed in the instantapplication are also useful in the treatment of these angiogenesisrelated cancers.

Tumors which have undergone neovascularization show an increasedpotential for metastasis. In fact, angiogenesis is essential for tumorgrowth and metastasis. (S. P. Cunningham, et al., Can. Research, 61:3206-3211 (2001)). The Akt inhibitors disclosed in the presentapplication are therefore also useful to prevent or decrease tumor cellmetastasis.

Further included within the scope of the invention is a method oftreating or preventing a disease in which angiogenesis is implicated,which is comprised of administering to a mammal in need of suchtreatment a therapeutically effective amount of a compound of thepresent invention. Ocular neovascular diseases are an example ofconditions where much of the resulting tissue damage can be attributedto aberrant infiltration of blood vessels in the eye (see WO 00/30651,published 2 Jun. 2000). The undesirable infiltration can be triggered byischemic retinopathy, such as that resulting from diabetic retinopathy,retinopathy of prematurity, retinal vein occlusions, etc., or bydegenerative diseases, such as the choroidal neovascularization observedin age-related macular degeneration. Inhibiting the growth of bloodvessels by administration of the present compounds would thereforeprevent the infiltration of blood vessels and prevent or treat diseaseswhere angiogenesis is implicated, such as ocular diseases like retinalvascularization, diabetic retinopathy, age-related macular degeneration,and the like.

Further included within the scope of the invention is a method oftreating or preventing a non-malignant disease in which angiogenesis isimplicated, including but not limited to: ocular diseases (such as,retinal vascularization, diabetic retinopathy and age-related maculardegeneration), atherosclerosis, arthritis, psoriasis, obesity andAlzheimer's disease (Dredge et al., Expert Opin. Biol. Ther. (2002)2(8):953-966). In another embodiment, a method of treating or preventinga disease in which angiogenesis is implicated includes: ocular diseases(such as, retinal vascularization, diabetic retinopathy and age-relatedmacular degeneration), atherosclerosis, arthritis and psoriasis.

Further included within the scope of the invention is a method oftreating hyperproliferative disorders such as restenosis, inflammation,autoimmune diseases and allergy/asthma.

Further included within the scope of the instant invention is the use ofthe instant compounds to coat stents and therefore the use of theinstant compounds on coated stents for the treatment and/or preventionof restenosis (WO03/032809).

Further included within the scope of the instant invention is the use ofthe instant compounds for the treatment and/or prevention ofosteoarthritis (WO03/035048).

Further included within the scope of the invention is a method oftreating hyperinsulinism.

The compounds of the invention are also useful in preparing a medicamentthat is useful in treating the diseases described above, in particularcancer.

In an embodiment of the invention, the instant compound is a selectiveinhibitor whose inhibitory efficacy is dependent on the PH domain. Inthis embodiment, the compound exhibits a decrease in in vitro inhibitoryactivity or no in vitro inhibitory activity against truncated Aktproteins lacking the PH domain.

In a further embodiment, the instant compound is selected from the groupof a selective inhibitor of Akt1, a selective inhibitor of Akt2 and aselective inhibitor of both Akt1 and Akt2, preferably a selectiveinhibitor of Akt1.

In another embodiment, the instant compound is selected from the groupof a selective inhibitor of Akt1, a selective inhibitor of Akt2, aselective inhibitor of Akt3 and a selective inhibitor of two of thethree Akt isoforms.

In another embodiment, the instant compound is a selective inhibitor ofall three Akt isoforms, but is not an inhibitor of one, two or all ofsuch Akt isoforms that have been modified to delete the PH domain, thehinge region or both the PH domain and the hinge region.

The present invention is further directed to a method of inhibiting Aktactivity, preferably Akt1 activity, which comprises administering to amammal in need thereof a pharmaceutically effective amount of theinstant compound.

The compounds of this invention may be administered to mammals,including humans, either alone or, in combination with pharmaceuticallyacceptable carriers, excipients or diluents, in a pharmaceuticalcomposition, according to standard pharmaceutical practice. Thecompounds can be administered orally or parenterally, including theintravenous, intramuscular, intraperitoneal, subcutaneous, rectal andtopical routes of administration.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions and suchcompositions may contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as calcium carbonate, sodium carbonate,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, microcrystalline cellulose, sodiumcrosscarmellose, corn starch, or alginic acid; binding agents, forexample starch, gelatin, polyvinyl-pyrrolidone or acacia, andlubricating agents, for example, magnesium stearate, stearic acid ortalc. The tablets may be uncoated or they may be coated by knowntechniques to mask the unpleasant taste of the drug or delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a watersoluble taste masking material such as hydroxypropylmethyl-cellulose orhydroxypropylcellulose, or a time delay material such as ethylcellulose, cellulose acetate buryrate may be employed.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with watersoluble carrier such as polyethylene glycol or an oil medium, forexample peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active material in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethylene-oxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose, saccharin or aspartame.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as butylated hydroxyanisol or alpha-tocopherol.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present. These compositions may be preserved by theaddition of an anti-oxidant such as ascorbic acid.

The pharmaceutical compositions of the invention may also be in the formof an oil-in-water emulsion. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring phosphatides, for example soy bean lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening, flavouring agents, preservatives and antioxidants.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative, flavoring and coloring agentsand antioxidant.

The pharmaceutical compositions may be in the form of sterile injectableaqueous solutions. Among the acceptable vehicles and solvents that maybe employed are water, Ringer's solution and isotonic sodium chloridesolution.

The sterile injectable preparation may also be a sterile injectableoil-in-water microemulsion where the active ingredient is dissolved inthe oily phase. For example, the active ingredient may be firstdissolved in a mixture of soybean oil and lecithin. The oil solutionthen introduced into a water and glycerol mixture and processed to forma microemulation.

The injectable solutions or microemulsions may be introduced into apatient's blood-stream by local bolus injection. Alternatively, it maybe advantageous to administer the solution or microemulsion in such away as to maintain a constant circulating concentration of the instantcompound. In order to maintain such a constant concentration, acontinuous intravenous delivery device may be utilized. An example ofsuch a device is the Deltec CADD-PLUS™ model 5400 intravenous pump.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension for intramuscular andsubcutaneous administration. This suspension may be formulated accordingto the known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example as a solution in 1,3-butane diol. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose any bland fixed oil may be employed includingsynthetic mono- or diglycerides. In addition, fatty acids such as oleicacid find use in the preparation of injectables.

Compounds of the present invention may also be administered in the fowlof suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials include cocoa butter, glycerinatedgelatin, hydrogenated vegetable oils, mixtures of polyethylene glycolsof various molecular weights and fatty acid esters of polyethyleneglycol.

For topical use, creams, ointments, jellies, solutions or suspensions,etc., containing the compound of the present invention are employed.(For purposes of this application, topical application shall includemouth washes and gargles.)

The compounds for the present invention can be administered inintranasal form via topical use of suitable intranasal vehicles anddelivery devices, or via transdermal routes, using those forms oftransdermal skin patches well known to those of ordinary skill in theart. To be administered in the form of a transdermal delivery system,the dosage administration will, of course, be continuous rather thanintermittent throughout the dosage regimen. Compounds of the presentinvention may also be delivered as a suppository employing bases such ascocoa butter, glycerinated gelatin, hydrogenated vegetable oils,mixtures of polyethylene glycols of various molecular weights and fattyacid esters of polyethylene glycol.

When a composition according to this invention is administered into ahuman subject, the daily dosage will normally be determined by theprescribing physician with the dosage generally varying according to theage, weight, and response of the individual patient, as well as theseverity of the patient's symptoms.

The dosage regimen utilizing the compounds of the instant invention canbe selected in accordance with a variety of factors including type,species, age, weight, sex and the type of cancer being treated; theseverity (i.e., stage) of the cancer to be treated; the route ofadministration; the renal and hepatic function of the patient; and theparticular compound or salt thereof employed. An ordinarily skilledphysician or veterinarian can readily determine and prescribe theeffective amount of the drug required to treat, for example, to prevent,inhibit (fully or partially) or arrest the progress of the disease. Forexample, compounds of the instant invention can be administered in atotal daily dose of up to 10,000 mg. Compounds of the instant inventioncan be administered once daily (QD), or divided into multiple dailydoses such as twice daily (BID), and three times daily (TID). Compoundsof the instant invention can be administered at a total daily dosage ofup to 10,000 mg, e.g., 2,000 mg, 3,000 mg, 4,000 mg, 6,000 mg, 8,000 mgor 10,000 mg, which can be administered in one daily dose or can bedivided into multiple daily doses as described above.

For example, compounds of the instant invention can be administered in atotal daily dose of up to 1,000 mg. Compounds of the instant inventioncan be administered once daily (QD), or divided into multiple dailydoses such as twice daily (BID), and three times daily (TID). Compoundsof the instant invention can be administered at a total daily dosage ofup to 1,000 mg, e.g., 200 mg, 300 mg, 400 mg, 600 mg, 800 mg or 1,000mg, which can be administered in one daily dose or can be divided intomultiple daily doses as described above.

In addition, the administration can be continuous, i.e., every day, orintermittently. The terms “intermittent” or “intermittently” as usedherein means stopping and starting at either regular or irregularintervals. For example, intermittent administration of a compound of theinstant invention may be administration one to six days per week or itmay mean administration in cycles (e.g. daily administration for two toeight consecutive weeks, then a rest period with no administration forup to one week) or it may mean administration on alternate days.

In addition, the compounds of the instant invention may be administeredaccording to any of the schedules described above, consecutively for afew weeks, followed by a rest period. For example, the compounds of theinstant invention may be administered according to any one of theschedules described above from two to eight weeks, followed by a restperiod of one week, or twice daily at a dose of 100-500 mg for three tofive days a week. In another particular embodiment, the compounds of theinstant invention may be administered three times daily for twoconsecutive weeks, followed by one week of rest.

Any one or more of the specific dosages and dosage schedules of thecompounds of the instant invention, may also be applicable to any one ormore of the therapeutic agents to be used in the combination treatment(hereinafter referred to as the “second therapeutic agent”).

Moreover, the specific dosage and dosage schedule of this secondtherapeutic agent can further vary, and the optimal dose, dosingschedule and route of administration will be determined based upon thespecific second therapeutic agent that is being used.

Of course, the route of administration of the compounds of the instantinvention is independent of the route of administration of the secondtherapeutic agent. In an embodiment, the administration for a compoundof the instant invention is oral administration. In another embodiment,the administration for a compound of the instant invention isintravenous administration. Thus, in accordance with these embodiments,a compound of the instant invention is administered orally orintravenously, and the second therapeutic agent can be administeredorally, parenterally, intraperitoneally, intravenously, intraarterially,transdermally, sublingually, intramuscularly, rectally, transbuccally,intranasally, liposomally, via inhalation, vaginally, intraocularly, vialocal delivery by catheter or stent, subcutaneously, intraadiposally,intraarticularly, intrathecally, or in a slow release dosage form.

In addition, a compound of the instant invention and second therapeuticagent may be administered by the same mode of administration, i.e. bothagents administered e.g. orally, by IV. However, it is also within thescope of the present invention to administer a compound of the instantinvention by one mode of administration, e.g. oral, and to administerthe second therapeutic agent by another mode of administration, e.g. IVor any other ones of the administration modes described hereinabove.

The first treatment procedure, administration of a compound of theinstant invention, can take place prior to the second treatmentprocedure, i.e., the second therapeutic agent, after the treatment withthe second therapeutic agent, at the same time as the treatment with thesecond therapeutic agent, or a combination thereof. For example, a totaltreatment period can be decided for a compound of the instant invention.The second therapeutic agent can be administered prior to onset oftreatment with a compound of the instant invention or followingtreatment with a compound of the instant invention. In addition,anti-cancer treatment can be administered during the period ofadministration of a compound of the instant invention but does not needto occur over the entire treatment period of a compound of the instantinvention.

The instant compounds are also useful in combination with therapeutic,chemotherapeutic and anti-cancer agents. Combinations of the presentlydisclosed compounds with therapeutic, chemotherapeutic and anti-canceragents are within the scope of the invention. Examples of such agentscan be found in Cancer Principles and Practice of Oncology by V. T.Devita and S. Hellman (editors), 6^(th) edition (Feb. 15, 2001),Lippincott Williams & Wilkins Publishers. A person of ordinary skill inthe art would be able to discern which combinations of agents would beuseful based on the particular characteristics of the drugs and thecancer involved. Such agents include the following: estrogen receptormodulators, androgen receptor modulators, retinoid receptor modulators,cytotoxic/cytostatic agents, antiproliferative agents, prenyl-proteintransferase inhibitors, HMG-CoA reductase inhibitors and otherangiogenesis inhibitors, HIV protease inhibitors, reverse transcriptaseinhibitors, inhibitors of cell proliferation and survival signaling,bisphosphonates, aromatase inhibitors, siRNA therapeutics, γ-secretaseinhibitors, agents that interfere with receptor tyrosine kinases (RTKs)and agents that interfere with cell cycle checkpoints. The instantcompounds are particularly useful when co-administered with radiationtherapy.

“Estrogen receptor modulators” refers to compounds that interfere withor inhibit the binding of estrogen to the receptor, regardless ofmechanism. Examples of estrogen receptor modulators include, but are notlimited to, tamoxifen, raloxifene, idoxifene, LY353381, LY117081,toremifene, fulvestrant,4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-piperidinyl)ethoxy]phenyl]-2H-1-benzopyran-3-yl]-phenyl-2,2-dimethylpropanoate,4,4′-dihydroxybenzophenone-2,4-dinitrophenyl-hydrazone, and SH646.

“Androgen receptor modulators” refers to compounds which interfere orinhibit the binding of androgens to the receptor, regardless ofmechanism. Examples of androgen receptor modulators include finasterideand other 5α-reductase inhibitors, nilutamide, flutamide, bicalutamide,liarozole, and abiraterone acetate.

“Retinoid receptor modulators” refers to compounds which interfere orinhibit the binding of retinoids to the receptor, regardless ofmechanism. Examples of such retinoid receptor modulators includebexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid,α-difluoromethylornithine, ILX23-7553, trans-N-(4′-hydroxyphenyl)retinamide, and N-4-carboxyphenyl retinamide.

“Cytotoxic/cytostatic agents” refer to compounds which cause cell deathor inhibit cell proliferation primarily by interfering directly with thecell's functioning or inhibit or interfere with cell myosis, includingalkylating agents, tumor necrosis factors, intercalators, hypoxiaactivatable compounds, microtubule inhibitors/microtubule-stabilizingagents, inhibitors of mitotic kinesins, histone deacetylase inhibitors,inhibitors of kinases involved in mitotic progression, inhibitors ofkinases involved in growth factor and cytokine signal transductionpathways, antimetabolites, biological response modifiers,hormonal/anti-hormonal therapeutic agents, haematopoietic growthfactors, monoclonal antibody targeted therapeutic agents, topoisomeraseinhibitors, proteosome inhibitors, ubiquitin ligase inhibitors, andaurora kinase inhibitors.

Examples of cytotoxic/cytostatic agents include, but are not limited to,sertenef, cachectin, ifosfamide, tasonermin, lonidamine, carboplatin,altretamine, prednimustine, dibromodulcitol, ranimustine, fotemustine,nedaplatin, oxaliplatin, temozolomide, heptaplatin, estramustine,improsulfan tosilate, trofosfamide, nimustine, dibrospidium chloride,pumitepa, lobaplatin, satraplatin, profiromycin, cisplatin, irofulven,dexifosfamide, cis-aminedichloro(2-methyl-pyridine)platinum,benzylguanine, glufosfamide, GPX100, (trans, trans,trans)-bis-mu-(hexane-1,6-diamine)-mu-[diamine-platinum(II)]bis[diamine(chloro)platinum(II)]tetrachloride, diarizidinylspermine, arsenic trioxide,1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin,idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin,pinafide, valrubicin, amrubicin, antineoplaston,3′-deamino-3′-morpholino-13-deoxo-10-hydroxycaminomycin, annamycin,galarubicin, elinafide, MEN10755,4-demethoxy-3-deamino-3-aziridinyl-4-methylsulphonyl-daunorubicin (seeWO 00/50032), Raf kinase inhibitors (such as Bay43-9006) and mTORinhibitors (such as Wyeth's CCI-779).

An example of a hypoxia activatable compound is tirapazamine.

Examples of proteosome inhibitors include but are not limited tolactacystin and MLN-341 (Velcade).

Examples of microtubule inhibitors/microtubule-stabilising agentsinclude paclitaxel, vindesine sulfate,3′,4′-didehydro-4′-deoxy-8′-norvincaleulcoblastine, docetaxol, rhizoxin,dolastatin, mivobulin isethionate, auristatin, cemadotin, RPR109881,BMS184476, vinflunine, cryptophycin,2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzene sulfonamide,anhydrovinblastine,N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t-butylamide,TDX258, the epothilones (see for example U.S. Pat. Nos. 6,284,781 and6,288,237) and BMS188797. In an embodiment the epothilones are notincluded in the microtubule inhibitors/microtubule-stabilising agents.

Some examples of topoisomerase inhibitors are topotecan, hycaptamine,irinotecan, rubitecan,6-ethoxypropionyl-3′,4′-O-exo-benzylidene-chartreusin,9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-kl]acridine-2-(6H)propanamine,1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3′,4′:b,7]-indolizino[1,2b]quinoline-10,13(9H,15H)dione,lurtotecan, 7-[2-(N-isopropylamino)ethyl]-(20S)camptothecin, BNP1350,BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane,2′-dimethylamino-2′-deoxy-etoposide, GL331,N-[2-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazole-1-carboxamide,asulacrine, (5a, 5aB,8aa,9b)-9-[2-[N-[2-(dimethylamino)ethyl]-N-methylamino]ethyl]-5-[4-hydroxy-3,5-dimethoxyphenyl]-5,5a,6,8,8a,9-hexohydrofuro(3′,4′:6,7)naphtho(2,3-d)-1,3-dioxol-6-one,2,3-(methylenedioxy)-5-methyl-7-hydroxy-8-methoxybenzo[c]-phenanthridinium,6,9-bis[(2-aminoethyl)amino]benzo[g]isoquinoline-5,10-dione,5-(3-aminopropylamino)-7,10-dihydroxy-2-(2-hydroxyethylaminomethyl)-6H-pyrazolo[4,5,1-de]acridin-6-one,N-[1-[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl]formamide,N-(2-(dimethylamino)ethyl)acridine-4-carboxamide,6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno[2,1-e]quinolin-7-one,and dimesna.

Examples of inhibitors of mitotic kinesins, and in particular the humanmitotic kinesin KSP, are described in Publications WO03/039460,WO03/050064, WO03/050122, WO03/049527, WO03/049679, WO03/049678,WO04/039774, WO03/079973, WO03/099211, WO03/105855, WO03/106417,WO04/037171, WO04/058148, WO04/058700, WO04/126699, WO05/018638,WO05/019206, WO05/019205, WO05/018547, WO05/017190, US200510176776. Inan embodiment inhibitors of mitotic kinesins include, but are notlimited to inhibitors of KSP, inhibitors of MKLP1, inhibitors of CENP-E,inhibitors of MCAK and inhibitors of Rab6-KIFL.

Examples of “histone deacetylase inhibitors” include, but are notlimited to, SAHA, TSA, oxamflatin, PXD101, MG98 and scriptaid. Furtherreference to other histone deacetylase inhibitors may be found in thefollowing manuscript; Miller, T. A. et al. J. Med. Chem.46(24):5097-5116 (2003).

“Inhibitors of kinases involved in mitotic progression” include, but arenot limited to, inhibitors of aurora kinase, inhibitors of Polo-likekinases (PLK; in particular inhibitors of PLK-1), inhibitors of bub-1and inhibitors of bub-R1. An example of an “aurora kinase inhibitor” isVX-680.

“Antiproliferative agents” includes antisense RNA and DNAoligonucleotides such as G3139, ODN698, RVASKRAS, GEM231, and INX3001,and antimetabolites such as enocitabine, carmofur, tegafur, pentostatin,doxifluridine, trimetrexate, fludarabine, capecitabine, galocitabine,cytarabine ocfosfate, fosteabine sodium hydrate, raltitrexed,paltitrexid, emitefur, tiazofurin, decitabine, nolatrexed, pemetrexed,nelzarabine, 2′-deoxy-2′-methylidenecytidine,2′-fluoromethylene-2′-deoxycytidine,N-[5-(2,3-dihydro-benzofuryl)sulfonyl]-N′-(3,4-dichlorophenyl)urea,N6-[4-deoxy-4-[N2-[2-(E),4(E)-tetradecadienoyl]glycylamino]-L-glycero-B-L-manno-heptopyranosyl]adenine,aplidine, ecteinascidin, troxacitabine,4-[2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b][1,4]thiazin-6-yl-(S)-ethyl]-2,5-thienoyl-L-glutamicacid, aminopterin, 5-fluorouracil, alanosine,11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6-methoxy-14-oxa-1,11-diazatetracyclo(7.4.1.0.0)-tetradeca-2,4,6-trien-9-ylacetic acid ester, swainsonine, lometrexol, dexrazoxane, methioninase,2′-cyano-2′-deoxy-N4-palmitoyl-1-B-D-arabino furanosyl cytosine,3-aminopyridine-2-carboxaldehyde thiosemicarbazone and trastuzumab.

Examples of monoclonal antibody targeted therapeutic agents includethose therapeutic agents which have cytotoxic agents or radioisotopesattached to a cancer cell specific or target cell specific monoclonalantibody. Examples include Bexxar.

“HMG-CoA reductase inhibitors” refers to inhibitors of3-hydroxy-3-methylglutaryl-CoA reductase. Examples of HMG-CoA reductaseinhibitors that may be used include but are not limited to lovastatin(MEVACOR®; see U.S. Pat. Nos. 4,231,938, 4,294,926 and 4,319,039),simvastatin (ZOCOR®; see U.S. Pat. Nos. 4,444,784, 4,820,850 and4,916,239), pravastatin (PRAVACHOL®; see U.S. Pat. Nos. 4,346,227,4,537,859, 4,410,629, 5,030,447 and 5,180,589), fluvastatin (LESCOL®;see U.S. Pat. Nos. 5,354,772, 4,911,165, 4,929,437, 5,189,164,5,118,853, 5,290,946 and 5,356,896), atorvastatin (LIPITOR®; see U.S.Pat. Nos. 5,273,995, 4,681,893, 5,489,691 and 5,342,952) andcerivastatin (also known as rivastatin and BAYCHOL®; see U.S. Pat. No.5,177,080). The structural formulas of these and additional HMG-CoAreductase inhibitors that may be used in the instant methods aredescribed at page 87 of M. Yalpani, “Cholesterol Lowering Drugs”,Chemistry & Industry, pp. 85-89 (5 Feb. 1996) and U.S. Pat. Nos.4,782,084 and 4,885,314. The term HMG-CoA reductase inhibitor as usedherein includes all pharmaceutically acceptable lactone and open-acidforms (i.e., where the lactone ring is opened to form the free acid) aswell as salt and ester forms of compounds which have HMG-CoA reductaseinhibitory activity, and therefor the use of such salts, esters,open-acid and lactone forms is included within the scope of thisinvention.

“Prenyl-protein transferase inhibitor” refers to a compound whichinhibits any one or any combination of the prenyl-protein transferaseenzymes, including farnesyl-protein transferase (FPTase),geranylgeranyl-protein transferase type I (GGPTase-I), andgeranylgeranyl-protein transferase type-II (GGPTase-II, also called RabGGPTase).

Examples of prenyl-protein transferase inhibitors can be found in thefollowing publications and patents: WO 96/30343, WO 97/18813, WO97/21701, WO 97/23478, WO 97/38665, WO 98/28980, WO 98/29119, WO95/32987, U.S. Pat. No. 5,420,245, U.S. Pat. No. 5,523,430, U.S. Pat.No. 5,532,359, U.S. Pat. No. 5,510,510, U.S. Pat. No. 5,589,485, U.S.Pat. No. 5,602,098, European Patent Publ. 0 618 221, European PatentPubl. 0 675 112, European Patent Publ. 0 604 181, European Patent Publ.0 696 593, WO 94/19357, WO 95/08542, WO 95/11917, WO 95/12612, WO95/12572, WO 95/10514, U.S. Pat. No. 5,661,152, WO 95/10515, WO95/10516, WO 95/24612, WO 95/34535, WO 95/25086, WO 96/05529, WO96/06138, WO 96/06193, WO 96/16443, WO 96/21701, WO 96/21456, WO96/22278, WO 96/24611, WO 96/24612, WO 96/05168, WO 96/05169, WO96/00736, U.S. Pat. No. 5,571,792, WO 96/17861, WO 96/33159, WO96/34850, WO 96/34851, WO 96/30017, WO 96/30018, WO 96/30362, WO96/30363, WO 96/31111, WO 96/31477, WO 96/31478, WO 96/31501, WO97/00252, WO 97/03047, WO 97/03050, WO 97/04785, WO 97/02920, WO97/17070, WO 97/23478, WO 97/26246, WO 97/30053, WO 97/44350, WO98/02436, and U.S. Pat. No. 5,532,359. For an example of the role of aprenyl-protein transferase inhibitor on angiogenesis see European J. ofCancer, Vol. 35, No. 9, pp. 1394-1401 (1999).

“Angiogenesis inhibitors” refers to compounds that inhibit the formationof new blood vessels, regardless of mechanism. Examples of angiogenesisinhibitors include, but are not limited to, tyrosine kinase inhibitors,such as inhibitors of the tyrosine kinase receptors Flt-1 (VEGFR1) andFlk-1/KDR (VEGFR2), inhibitors of epidermal-derived, fibroblast-derived,or platelet derived growth factors, MMP (matrix metalloprotease)inhibitors, integrin blockers, interferon-α, interleukin-12, pentosanpolysulfate, cyclooxygenase inhibitors, including nonsteroidalanti-inflammatories (NSAIDs) like aspirin and ibuprofen as well asselective cyclooxy-genase-2 inhibitors like celecoxib and rofecoxib(PNAS, Vol. 89, p. 7384 (1992); JNCI, Vol. 69, p. 475 (1982); Arch.Opthalmol., Vol. 108, p. 573 (1990); Anat. Rec., Vol. 238, p. 68 (1994);FEBS Letters, Vol. 372, p. 83 (1995); Clin, Orthop. Vol. 313, p. 76(1995); J. Mol. Endocrinol., Vol. 16, p. 107 (1996); Jpn. J. Pharmacol.,Vol. 75, p. 105 (1997); Cancer Res., Vol. 57, p. 1625 (1997); Cell, Vol.93, p. 705 (1998); Intl. J. Mol. Med., Vol. 2, p. 715 (1998); J. Biol.Chem., Vol. 274, p. 9116 (1999)), steroidal anti-inflammatories (such ascorticosteroids, mineralocorticoids, dexamethasone, prednisone,prednisolone, methylpred, betamethasone), carboxyamidotriazole,combretastatin A-4, squalamine, 6-O-chloroacetyl-carbonyl)-fumagillol,thalidomide, angiostatin, troponin-1, angiotensin II antagonists (seeFernandez et al., J. Lab. Clin. Med. 105:141-145 (1985)), and antibodiesto VEGF (see, Nature Biotechnology, Vol. 17, pp. 963-968 (October 1999);Kim et al., Nature, 362, 841-844 (1993); WO 00/44777; and WO 00/61186).

Other therapeutic agents that modulate or inhibit angiogenesis and mayalso be used in combination with the compounds of the instant inventioninclude agents that modulate or inhibit the coagulation and fibrinolysissystems (see review in Clin. Chem. La. Med. 38:679-692 (2000)). Examplesof such agents that modulate or inhibit the coagulation and fibrinolysispathways include, but are not limited to, heparin (see Thromb. Haemost.80:10-23 (1998)), low molecular weight heparins and carboxypeptidase Uinhibitors (also known as inhibitors of active thrombin activatablefibrinolysis inhibitor [TAFIa]) (see Thrombosis Res. 101:329-354(2001)). TAM inhibitors have been described in U.S. Ser. Nos. 60/310,927(filed Aug. 8, 2001) and 60/349,925 (filed Jan. 18, 2002).

“Agents that interfere with cell cycle checkpoints” refer to compoundsthat inhibit protein kinases that transduce cell cycle checkpointsignals, thereby sensitizing the cancer cell to DNA damaging agents.Such agents include inhibitors of ATR, ATM, the CHK11 and CHK12 kinasesand cdk and cdc kinase inhibitors and are specifically exemplified by7-hydroxystaurosporin, flavopiridol, CYC202 (Cyclacel) and BMS-387032.

“Agents that interfere with receptor tyrosine kinases (RTKs)” refer tocompounds that inhibit RTKs and therefore mechanisms involved inoncogenesis and tumor progression. Such agents include inhibitors ofc-Kit, Eph, PDGF, Flt3 and c-Met. Further agents include inhibitors ofRTKs as described by Bume-Jensen and Hunter, Nature, 411:355-365, 2001.

“Inhibitors of cell proliferation and survival signalling pathway” referto compounds that inhibit signal transduction cascades downstream ofcell surface receptors. Such agents include inhibitors ofserine/threonine kinases (including but not limited to inhibitors of Aktsuch as described in WO 02/083064, WO 02/083139, WO 02/083140, US2004-0116432, WO 02/083138, US 2004-0102360, WO 03/086404, WO 03/086279,WO 03/086394, WO 03/084473, WO 03/086403, WO 2004/041162, WO2004/096131, WO 2004/096129, WO 2004/096135, WO 2004/096130, WO2005/100356, WO 2005/100344, US 2005/029941, US 2005/44294, US2005/43361, 60/734,188, 60/652,737, 60/670,469), inhibitors of Rafkinase (for example BAY-43-9006), inhibitors of MEK (for example CI-1040and PD-098059), inhibitors of mTOR (for example Wyeth CCI-779), andinhibitors of PI3K (for example LY294002).

As described above, the combinations with NSAID's are directed to theuse of NSAID's which are potent COX-2 inhibiting agents. For purposes ofthis specification an NSAID is potent if it possesses an IC₅₀ for theinhibition of COX-2 of 1 μM or less as measured by cell or microsomalassays.

The invention also encompasses combinations with NSAID's which areselective COX-2 inhibitors. For purposes of this specification NSAID'swhich are selective inhibitors of COX-2 are defined as those whichpossess a specificity for inhibiting COX-2 over COX-1 of at least 100fold as measured by the ratio of IC₅₀ for COX-2 over IC₅₀ for COX-1evaluated by cell or microsomal assays. Such compounds include, but arenot limited to those disclosed in U.S. Pat. No. 5,474,995, U.S. Pat. No.5,861,419, U.S. Pat. No. 6,001,843, U.S. Pat. No. 6,020,343, U.S. Pat.No. 5,409,944, U.S. Pat. No. 5,436,265, U.S. Pat. No. 5,536,752, U.S.Pat. No. 5,550,142, U.S. Pat. No. 5,604,260, U.S. Pat. No. 5,698,584,U.S. Pat. No. 5,710,140, WO 94/15932, U.S. Pat. No. 5,344,991, U.S. Pat.No. 5,134,142, U.S. Pat. No. 5,380,738, U.S. Pat. No. 5,393,790, U.S.Pat. No. 5,466,823, U.S. Pat. No. 5,633,272 and U.S. Pat. No. 5,932,598,all of which are hereby incorporated by reference.

Inhibitors of COX-2 that are particularly useful in the instant methodof treatment are: 3-phenyl-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone;and5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-methyl-5-pyridinyl)pyridine; ora pharmaceutically acceptable salt thereof.

Compounds that have been described as specific inhibitors of COX-2 andare therefore useful in the present invention include, but are notlimited to, the following: parecoxib, BEXTRA® and CELEBREX® or apharmaceutically acceptable salt thereof.

Other examples of angiogenesis inhibitors include, but are not limitedto, endostatin, ukrain, ranpirnase, IM862,5-methoxy-4-[2-methyl-3-(3-methyl-2-butenyl)oxiranyl]-1-oxaspiro[2,5]oct-6-yl(chloroacetyl)carbamate,acetyldinanaline,5-amino-1-[[3,5-dichloro-4-(4-chlorobenzoyl)phenyl]methyl]-1H-1,2,3-triazole-4-carboxamide,CM101, squalamine, combretastatin, RPI4610, NX31838, sulfatedmannopentaose phosphate,7,7-(carbonyl-bis[imino-N-methyl-4,2-pyrrolocarbonylimino[N-methyl-4,2-pyrrole]-carbonylimino]-bis-(1,3-naphthalenedisulfonate), and 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone(SU5416).

As used above, “integrin blockers” refers to compounds which selectivelyantagonize, inhibit or counteract binding of a physiological ligand tothe α_(v)β₃ integrin, to compounds which selectively antagonize, inhibitor counteract binding of a physiological ligand to the αvβ5 integrin, tocompounds which antagonize, inhibit or counteract binding of aphysiological ligand to both the α_(v)β₃ integrin and the α_(v)β₅integrin, and to compounds which antagonize, inhibit or counteract theactivity of the particular integrin(s) expressed on capillaryendothelial cells. The term also refers to antagonists of the α_(v)β₆,α_(v)β₈, α₅β₁, α₆β₁ and α₆β₄ integrins. The term also refers toantagonists of any combination of α_(v)β₃, α_(v)β₅, α_(v)β₆, α_(v)β₈,α₁β₁, α₂β₁, α₅β₁, α₆β₁ and α₆β₄ integrins.

Some specific examples of tyrosine kinase inhibitors includeN-(trifluoromethylphenyl)-5-methylisoxazol-4-carboxamide,3-[(2,4-dimethylpyrrol-5-yl)methylidenyl)indolin-2-one,17-(allylamino)-17-demethoxygeldanamycin,4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-[3-(4-morpholinyl)propoxyl]quinazoline,N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine,BIBX1382,2,3,9,10,11,12-hexahydro-10-(hydroxymethyl)-10-hydroxy-9-methyl-9,12-epoxy-1H-diindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocin-1-one,SH268, genistein, ST1571, CEP2563,4-(3-chlorophenylamino)-5,6-dimethyl-7H-pyrrolo[2,3-d]pyrimidinemethanesulfonate, 4-(3-bromo-4-hydroxyphenyl)amino-6,7-dimethoxyquinazoline,4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, SU6668, STI571A,N-4-chlorophenyl-4-(4-pyridylmethyl)-1-phthalazinamine, CI-1033, CDP860,ZR6474, RTK-787, CP549632, and CT53518.

Combinations with compounds other than anti-cancer compounds are alsoencompassed in the instant methods. For example, combinations of theinstantly claimed compounds with PPAR-γ (i.e., PPAR-gamma) agonists andPPAR-δ (i.e., PPAR-delta) agonists are useful in the treatment ofcertain malignancies. PPAR-γ and PPAR-δ are the nuclear peroxisomeproliferator-activated receptors γ and δ. The expression of PPAR-γ onendothelial cells and its involvement in angiogenesis has been reportedin the literature (see J. Cardiovasc. Pharmacol. 1998; 31:909-913; J.Biol. Chem. 1999; 274:9116-9121; Invest. Ophthalmol. Vis. Sci. 2000;41:2309-2317). More recently, PPAR-γ agonists have been shown to inhibitthe angiogenic response to VEGF in vitro; both troglitazone androsiglitazone maleate inhibit the development of retinalneovascularization in mice. (Arch. Ophthamol. 2001; 119:709-717).Examples of PPAR-γ agonists and PPAR-γ/α agonists include, but are notlimited to, thiazolidinediones (such as DRF2725, CS-011, troglitazone,rosiglitazone, and pioglitazone), fenofibrate, gemfibrozil, clofibrate,GW2570, SB219994, AR-H039242, JTT-501, MCC-555, GW2331, GW409544,NN2344, KRP297, NP0110, DRF4158, NN622, G1262570, PNU182716, DRF552926,2-[(5,7-dipropyl-3-trifluoromethyl-1,2-benzisoxazol-6-yl)oxy]-2-methylpropionicacid (disclosed in U.S. Ser. No. 09/782,856), and2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylicacid (disclosed in U.S. Ser. No. 60/235,708 and 60/244,697).

Another embodiment of the instant invention is the use of the presentlydisclosed compounds in combination with gene therapy for the treatmentof cancer. For an overview of genetic strategies to treating cancer seeHall et al (Am. J. Hum. Genet. 61:785-789, 1997) and Kufe et al (CancerMedicine, 5th Ed, pp 876-889, BC Decker, Hamilton 2000). Gene therapycan be used to deliver any tumor suppressing gene. Examples of suchgenes include, but are not limited to, p53, which can be delivered viarecombinant virus-mediated gene transfer (see U.S. Pat. No. 6,069,134,for example), a uPA/uPAR antagonist (“Adenovirus-Mediated Delivery of auPA/uPAR Antagonist Suppresses Angiogenesis-Dependent Tumor Growth andDissemination in Mice,” Gene Therapy, August 1998; 5(8):1105-13), andinterferon gamma (J. Immunol. 2000; 164:217-222).

The compounds of the instant invention may also be administered incombination with an inhibitor of inherent multidrug resistance (MDR), inparticular MDR associated with high levels of expression of transporterproteins. Such MDR inhibitors include inhibitors of p-glycoprotein(P-gp), such as LY335979, XR9576, OC144-093, R101922, VX853 and PSC833(valspodar).

A compound of the present invention may be employed in conjunction withanti-emetic agents to treat nausea or emesis, including acute, delayed,late-phase, and anticipatory emesis, which may result from the use of acompound of the present invention, alone or with radiation therapy. Forthe prevention or treatment of emesis, a compound of the presentinvention may be used in conjunction with other anti-emetic agents,especially neurokinin-1 receptor antagonists, 5HT3 receptor antagonists,such as ondansetron, granisetron, tropisetron, and zatisetron, GABABreceptor agonists, such as baclofen, a corticosteroid such as Decadron(dexamethasone), Kenalog, Aristocort, Nasalide, Preferid, Benecorten orothers such as disclosed in U.S. Pat. Nos. 2,789,118, 2,990,401,3,048,581, 3,126,375, 3,929,768, 3,996,359, 3,928,326 and 3,749,712, anantidopaminergic, such as the phenothiazines (for exampleprochlorperazine, fluphenazine, thioridazine and mesoridazine),metoclopramide or dronabinol. In another embodiment, conjunctive therapywith an anti-emesis agent selected from a neurokinin-1 receptorantagonist, a 5HT3 receptor antagonist and a corticosteroid is disclosedfor the treatment or prevention of emesis that may result uponadministration of the instant compounds.

Neurokinin-1 receptor antagonists of use in conjunction with thecompounds of the present invention are fully described, for example, inU.S. Pat. Nos. 5,162,339, 5,232,929, 5,242,930, 5,373,003, 5,387,595,5,459,270, 5,494,926, 5,496,833, 5,637,699, 5,719,147; European PatentPublication Nos. EP 0 360 390, 0 394 989, 0 428 434, 0 429 366, 0 430771, 0 436 334, 0 443 132, 0 482 539, 0 498 069, 0 499 313, 0 512 901, 0512 902, 0 514 273, 0 514 274, 0 514 275, 0 514 276, 0 515 681, 0 517589, 0 520 555, 0 522 808, 0 528 495, 0 532 456, 0 533 280, 0 536 817, 0545 478, 0 558 156, 0 577 394, 0 585 913, 0 590 152, 0 599 538, 0 610793, 0 634 402, 0 686 629, 0 693 489, 0 694 535, 0 699 655, 0 699 674, 0707 006, 0 708 101, 0 709 375, 0 709 376, 0 714 891, 0 723 959, 0 733632 and 0 776 893; PCT International Patent Publication Nos. WO90/05525, 90/05729, 91/09844, 91/18899, 92/01688, 92/06079, 92/12151,92/15585, 92/17449, 92/20661, 92/20676, 92/21677, 92/22569, 93/00330,93/00331, 93/01159, 93/01165, 93/01169, 93/01170, 93/06099, 93/09116,93/10073, 93/14084, 93/14113, 93/18023, 93/19064, 93/21155, 93/21181,93/23380, 93/24465, 94/00440, 94/01402, 94/02461, 94/02595, 94/03429,94/03445, 94/04494, 94/04496, 94/05625, 94/07843, 94/08997, 94/10165,94/10167, 94/10168, 94/10170, 94/11368, 94/13639, 94/13663, 94/14767,94/15903, 94/19320, 94/19323, 94/20500, 94/26735, 94/26740, 94/29309,95/02595, 95/04040, 95/04042, 95/06645, 95/07886, 95/07908, 95/08549,95/11880, 95/14017, 95/15311, 95/16679, 95/17382, 95/18124, 95/18129,95/19344, 95/20575, 95/21819, 95/22525, 95/23798, 95/26338, 95/28418,95/30674, 95/30687, 95/33744, 96/05181, 96/05193, 96/05203, 96/06094,96/07649, 96/10562, 96/16939, 96/18643, 96/20197, 96/21661, 96/29304,96/29317, 96/29326, 96/29328, 96/31214, 96/32385, 96/37489, 97/01553,97/01554, 97/03066, 97/08144, 97/14671, 97/17362, 97/18206, 97/19084,97/19942 and 97/21702; and in British Patent Publication Nos. 2 266 529,2 268 931, 2 269 170, 2 269 590, 2 271 774, 2 292 144, 2 293 168, 2 293169, and 2 302 689. The preparation of such compounds is fully describedin the aforementioned patents and publications, which are incorporatedherein by reference.

In an embodiment, the neurokinin-1 receptor antagonist for use inconjunction with the compounds of the present invention is selectedfrom:2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine,or a pharmaceutically acceptable salt thereof, which is described inU.S. Pat. No. 5,719,147.

A compound of the instant invention may also be administered with anagent useful in the treatment of anemia. Such an anemia treatment agentis, for example, a continuous eythropoiesis receptor activator (such asepoetin alfa).

A compound of the instant invention may also be administered with anagent useful in the treatment of neutropenia. Such a neutropeniatreatment agent is, for example, a hematopoietic growth factor whichregulates the production and function of neutrophils such as a humangranulocyte colony stimulating factor, (G-CSF). Examples of a G-CSFinclude filgrastim.

A compound of the instant invention may also be administered with animmunologic-enhancing drug, such as levamisole, isoprinosine andZadaxin.

A compound of the instant invention may also be useful for treating orpreventing cancer in combination with P450 inhibitors including:xenobiotics, quinidine, tyramine, ketoconazole, testosterone, quinine,methyrapone, caffeine, phenelzine, doxorubicin, troleandomycin,cyclobenzaprine, erythromycin, cocaine, furafyline, cimetidine,dextromethorphan, ritonavir, indinavir, amprenavir, diltiazem,terfenadine, verapamil, cortisol, itraconazole, mibefradil, nefazodoneand nelfinavir.

A compound of the instant invention may also be useful for treating orpreventing cancer in combination with Pgp and/or BCRP inhibitorsincluding: cyclosporin A, PSC833, GF120918, cremophorEL, fumitremorginC, Ko132, Ko134, Iressa, Imatnib mesylate, EKI-785, Cl1033, novobiocin,diethylstilbestrol, tamoxifen, resperpine, VX-710, tryprostatin A,flavonoids, ritonavir, saquinavir, nelfinavir, omeprazole, quinidine,verapamil, terfenadine, ketoconazole, nifidepine, FK506, amiodarone,XR9576, indinavir, amprenavir, cortisol, testosterone, LY335979,OC144-093, erythromycin, vincristine, digoxin and talinolol.

A compound of the instant invention may also be useful for treating orpreventing cancer, including bone cancer, in combination withbisphosphonates (understood to include bisphosphonates, diphosphonates,bisphosphonic acids and diphosphonic acids). Examples of bisphosphonatesinclude but are not limited to: etidronate (Didronel), pamidronate(Aredia), alendronate (Fosamax), risedronate (Actonel), zoledronate(Zometa), ibandronate (Boniva), incadronate or cimadronate, clodronate,EB-1053, minodronate, neridronate, piridronate and tiludronate includingany and all pharmaceutically acceptable salts, derivatives, hydrates andmixtures thereof.

A compound of the instant invention may also be useful for treating orpreventing breast cancer in combination with aromatase inhibitors.Examples of aromatase inhibitors include but are not limited to:anastrozole, letrozole and exemestane.

A compound of the instant invention may also be useful for treating orpreventing cancer in combination with siRNA therapeutics.

The compounds of the instant invention may also be administered incombination with γ-secretase inhibitors and/or inhibitors of NOTCHsignaling. Such inhibitors include compounds described in WO 01/90084,WO 02/30912, WO 01/70677, WO 03/013506, WO 02/36555, WO 03/093252, WO03/093264, WO 03/093251, WO 03/093253, WO 2004/039800, WO 2004/039370,WO 2005/030731, WO 2005/014553, U.S. Ser. No. 10/957,251, WO2004/089911, WO 02/081435, WO 02/081433, WO 03/018543, WO 2004/031137,WO 2004/031139, WO 2004/031138, WO 2004/101538, WO 2004/101539 and WO02/47671 (including LY-450139).

Inhibitors of Akt, as disclosed in the following publications; WO02/083064, WO 02/083139, WO 02/083140, US 2004-0116432, WO 02/083138, US2004-0102360, WO 03/086404, WO 03/086279, WO 03/086394, WO 03/084473, WO03/086403, WO 2004/041162, WO 2004/096131, WO 2004/096129, WO2004/096135, WO 2004/096130, WO 2005/100356, WO 2005/100344, US2005/029941, US 2005/44294, US 2005/43361, 60/734,188, 60/652,737,60/670,469, and including compounds of the instant invention, are alsouseful in combination with potassium salts, magnesium salts,beta-blockers (such as atenolol) and endothelin-a (ETa) antagonists withthe goal of maintaining cardiovascular homeostasis.

Inhibitors of Akt, as disclosed in the following publications; WO02/083064, WO 02/083139, WO 02/083140, US 2004-0116432, WO 02/083138, US2004-0102360, WO 03/086404, WO 03/086279, WO 03/086394, WO 03/084473, WO03/086403, WO 2004/041162, WO 2004/096131, WO 2004/096129, WO2004/096135, WO 2004/096130, WO 2005/100356, WO 2005/100344, US2005/029941, US 2005/44294, US 2005/43361, 60/734,188, 60/652,737,60/670,469, and including compounds of the instant invention, are alsouseful in combination with insulin, insulin secretagogues, PPAR-gammaagonists, metformin, somatostatin receptor agonists such as octreotide,DPP4 inhibitors, sulfonylureas and alpha-glucosidase inhibitors with thegoal of maintaining glucose homeostasis.

A compound of the instant invention may also be useful for treating orpreventing cancer in combination with PARP inhibitors.

A compound of the instant invention may also be useful for treatingcancer in combination with the following therapeutic agents: abarelix(Plenaxis Depot®); (Actiq®); aldesleukin (Prokine®); Aldesleukin(Proleukin®); Alemtuzumab (Campath®); alfuzosin HCl (UroXatral®);alitretinoin (Panretin®); allopurinol (Zyloprim®); altretamine(Hexylen®); amifostine (Ethyol®); anastrozole (Arimidex®); (Anzemet®);(Anexsia®); aprepitant (Emend®); arsenic trioxide (Trisenox®);asparaginase (Elspar®); azacitidine (Vidaza®); bendamustinehydrochloride (Treanda®); bevacuzimab (Avastin®); bexarotene capsules(Targretin®); bexarotene gel (Targeting); bleornycin (Blenoxane®);bortezomib (Velcade®); (Brofenac®); busulfan intravenous (Busulflex®);busulfan oral (Myleran®); calusterone (Methosarb®); capecitabine(Xeloda®); carboplatin (Paraplatin®); cannustine (BCNU®, BiCNU®);carmustine (Gliadel®); carmustine with Polifeprosan 20 Implant (GliadelWafer celecoxib (Celebrex®); cetuximab (Erbitux®); chlorambucil(Leukeran®); cinacalcet (Sensipar®); cisplatin (Platinol®); cladribine(Leustatin®, 2-CdA®); clofarabine (Clolar®); cyclophosphamide (Cytoxan®,Neosar®); cyclophosphamide (Cytoxan Injection®); cyclophosphamide(Cytoxan Tablet®); cytarabine (Cytosar-U®); cytarabine liposomal(DepoCyt®); dacarbazine (DTIC-Dome®); dactinomycin, actinomycin D(Cosmegen®); Darbepoetin alfa (Aranesp®); dasatinib (Sprycel®);daunorubicin liposomal (DanuoXome®); daunorubicin, daunomycin(Daunorubicin®); daunorubicin, daunomycin (Cerubidine®); decitabine(Dacogen®); Denileukin diftitox (Ontak®); dexrazoxane (Zinecard®);docetaxel (Taxotere®); doxorubicin (Adriamycin PFS®); doxorubicin(Adriamycin®, Rubex®); doxorubicin (Adriamycin PFS Injection®);doxorubicin liposomal (Doxil®); dromostanolone propionate(Dromostanolone®); dromostanolone propionate (Masterone Injection®);Elliott's B Solution (Elliott's B Solution®); epirubicin (Ellence®);Epoetin alfa (Epogen®); erlotinib (Tarceva®); estramustine (Emcyt®);etoposide phosphate (Etopophos®); etoposide, VP-16 (Vepesid®);exemestane (Aromasin®); fentanyl citrate (Fentora®); Filgrastim(Neupogen®); floxuridine (intraarterial) (FUDR®); fludarabine(Fludara®); fluorouracil, 5-FU (Adrucil®); flutamide (Eulexin®);fulvestrant (Faslodex®); gefitinib (Iressa®); gemcitabine (Gemzar®);gemtuzumab ozogamicin (Mylotarg®); goserelin acetate (Zoladex Implant®);goserelin acetate (Zoladex®); granisetron (Kytril Solution®); histrelinacetate (Histrelin Implant®); hydroxyurea (Hydrea®); IbritumomabTiuxetan (Zevalin®); idarubicin (Idamycin®); ifosfamide (IFEX®);imatinib mesylate (Gleevec®); interferon alfa 2a (Roferon A®);Interferon alfa-2b (Intron A®); irinotecan (Camptosar®); (Kadian®);ixabepilone (Ixempra®); lapatinib (Tykerb®); lenalidomide (Revlimid®);letrozole (Femara®); leucovorin (Wellcovorin®, Leucovorin®); LeuprolideAcetate (Eligard®); (Lupron Depot®); (Viadur®); levamisole (Ergamisol®);lomustine, CCNU (CeeBU®); meclorethamine, nitrogen mustard (Mustargen®);megestrol acetate (Megace®); melphalan, L-PAM (Alkeran®);mercaptopurine, 6-MP (Purinethol®); mesna (Mesnex®); mesna (MesnexTabs®); methotrexate (Methotrexate®); methoxsalen (Uvadex®); mitomycin C(Mutamycin®); mitomycin C (Mitozytrex®); mitotane (Lysodren®);mitoxantrone (Novantrone®); nandrolone phenpropionate (Durabolin-500);nelarabine (Arranon®); nilotinib hydrochloride monohydrate (Tasigna®);Nofetumomab (Verluma®); Oprelvekin (Neumega®); (Neupogen®); oxaliplatin(Eloxatin®); paclitaxel (Paxene®); paclitaxel (Taxol®); paclitaxelprotein-bound particles (Abraxane®); palifel win (Kepivance®);palonosetron (Aloxi®); pamidronate (Aredia®); panitumumab (Vectibix®);pegademase (Adagen (Pegademase Bovine)®); pegaspargase (Oncaspar®);Pegfilgrastim (Neulasta®); pemetrexed disodium (Alimta®); pentostatin(Nipent®); pipobroman (Vercyte®); plicarnycin, mithramycin (Mithracin®);porfimer sodium (Photofrin®); procarbazine (Matulane®); (Quadramet®);quadrivalent human papillomavirus (types 6, 11, 16, 18) recombinantvaccine (Gardasil®); quinacrine (Atabrine®); raloxifene hydrochloride(Evista®); Rasburicase (Elitek®); Rituximab (Rituxan®); sargramostim(Leukine®); Sargramostim (Prokine®); secretin (SecreFlo®); sorafenib(Nexavar®); streptozocin (Zanosar®); sunitinib maleate (Sutent®); talc(Sclerosol®); tamoxifen (Nolvadex®); temozolomide (Temodar®);temsirolimus (Torisel®); teniposide, VM-26 (Vumon®); (Temodar®);testolactone (Teslac®); thalidomide (Thalomid®); thioguanine, 6-TG(Thioguanine®); thiotepa (Thioplex®); topotecan (Hycamtin®); toremifene(Fareston®); Tositumomab (Bexxar®); Tositumomab/I-131 tositumomab(Bexxar®); Trastuzumab (Herceptin®); (Trelstar LA®); tretinoin, ATRA(Vesanoid®); triptorelin pamoate (Trelstar Depot®); (UltraJect®); UracilMustard (Uracil Mustard Capsules®); valrubicin (Valstar®); vinblastine(Velban®); vincristine (Oncovin®); vinorelbine (Navelbine®); vorinostat(Zolinza®); (Zofran ODT®); and zoledronate (Zometa®).

The compounds of the instant invention are useful for treating cancer incombination with taxanes.

The compounds of the instant invention are useful for treating cancer incombination with docetaxel (Taxotere®).

The compounds of the instant invention are useful for treating cancer incombination with vorinostat (Zolinza®).

The compounds of the instant invention are useful for treating cancer incombination with the aurora kinase inhibitor, MK-0457.

The compounds of the instant invention are useful for treating cancer incombination with the mTOR inhibitor, AP 23573.

The compounds of the instant invention are useful for treating cancer incombination with the IGF1R inhibitor, MK-0646.

The compounds of the instant invention are useful for treating cancer incombination with satraplatin.

The compounds of the instant invention are useful for treating cancer incombination with lapatinib (Tykerb®).

Thus, the scope of the instant invention encompasses the use of theinstantly claimed compounds in combination with a second compoundselected from: an estrogen receptor modulator, an androgen receptormodulator, a retinoid receptor modulator, a cytotoxic/cytostatic agent,an antiproliferative agent, a prenyl-protein transferase inhibitor, anHMG-CoA reductase inhibitor, an HIV protease inhibitor, a reversetranscriptase inhibitor, an angiogenesis inhibitor, PPAR-γ agonists,PPAR-δ agonists, an inhibitor of inherent multidrug resistance, ananti-emetic agent, an agent useful in the treatment of anemia, an agentuseful in the treatment of neutropenia, an immunologic-enhancing drug,an inhibitor of cell proliferation and survival signaling, abisphosphonate, an aromatase inhibitor, an siRNA therapeutic,γ-secretase inhibitors, agents that interfere with receptor tyrosinekinases (RTKs), an agent that interferes with a cell cycle checkpointand any of the therapeutic agents listed above.

The term “administration” and variants thereof (e.g., “administering” acompound) in reference to a compound of the invention means introducingthe compound or a prodrug of the compound into the system of the animalin need of treatment. When a compound of the invention or prodrugthereof is provided in combination with one or more other active agents(e.g., a cytotoxic agent, etc.), “administration” and its variants areeach understood to include concurrent and sequential introduction of thecompound or prodrug thereof and other agents.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

The term “therapeutically effective amount” as used herein means thatamount of active compound or pharmaceutical agent that elicits thebiological or medicinal response in a tissue, system, animal or humanthat is being sought by a researcher, veterinarian, medical doctor orother clinician.

The term “treating cancer” or “treatment of cancer” refers toadministration to a mammal afflicted with a cancerous condition andrefers to an effect that alleviates the cancerous condition by killingthe cancerous cells, but also to an effect that results in theinhibition of growth and/or metastasis of the cancer.

In an embodiment, the angiogenesis inhibitor to be used as the secondcompound is selected from a tyrosine kinase inhibitor, an inhibitor ofepidermal-derived growth factor, an inhibitor of fibroblast-derivedgrowth factor, an inhibitor of platelet derived growth factor, an MMP(matrix metalloprotease) inhibitor, an integrin blocker, interferon-α,interleukin-12, pentosan polysulfate, a cyclooxygenase inhibitor,carboxyamidotriazole, combretastatin A-4, squalamine,6-O-chloroacetyl-carbonyl)-fumagillol, thalidomide, angiostatin,troponin-1, or an antibody to VEGF. In an embodiment, the estrogenreceptor modulator is tamoxifen or raloxifene.

Also included in the scope of the claims is a method of treating cancerthat comprises administering a therapeutically effective amount of acompound of the instant invention in combination with radiation therapyand/or in combination with a second compound selected from: an estrogenreceptor modulator, an androgen receptor modulator, a retinoid receptormodulator, a cytotoxic/cytostatic agent, an antiproliferative agent, aprenyl-protein transferase inhibitor, an HMG-CoA reductase inhibitor, anHIV protease inhibitor, a reverse transcriptase inhibitor, anangiogenesis inhibitor, PPAR-γ agonists, PPAR-δ agonists, an inhibitorof inherent multidrug resistance, an anti-emetic agent, an agent usefulin the treatment of anemia, an agent useful in the treatment ofneutropenia, an immunologic-enhancing drug, an inhibitor of cellproliferation and survival signaling, a bisphosphonate, an aromataseinhibitor, an siRNA therapeutic, γ-secretase inhibitors, agents thatinterfere with receptor tyrosine kinases (RTKs), an agent thatinterferes with a cell cycle checkpoint and any of the therapeuticagents listed above.

And yet another embodiment of the invention is a method of treatingcancer that comprises administering a therapeutically effective amountof a compound of the instant invention in combination with paclitaxel ortrastuzumab.

The invention further encompasses a method of treating or preventingcancer that comprises administering a therapeutically effective amountof a compound of the instant invention in combination with a COX-2inhibitor.

The instant invention also includes a pharmaceutical composition usefulfor treating or preventing cancer that comprises a therapeuticallyeffective amount of a compound of the instant invention and a secondcompound selected from: an estrogen receptor modulator, an androgenreceptor modulator, a retinoid receptor modulator, acytotoxic/cytostatic agent, an antiproliferative agent, a prenyl-proteintransferase inhibitor, an HMG-CoA reductase inhibitor, an HIV proteaseinhibitor, a reverse transcriptase inhibitor, an angiogenesis inhibitor,a PPAR-γ agonist, a PPAR-δ agonist, an inhibitor of cell proliferationand survival signaling, a bisphosphonate, an aromatase inhibitor, ansiRNA therapeutic, γ-secretase inhibitors, agents that interfere withreceptor tyrosine kinases (RTKs), an agent that interferes with a cellcycle checkpoint and any of the therapeutic agents listed above.

All patents, publications and pending patent applications identified arehereby incorporated by reference.

Abbreviations used in the description of the chemistry and in theExamples that follow are: AEBSF (p-aminoethylbenzenesulfonyl fluoride);BSA (bovine serum albumin); BuLi (n-Butyl lithium); CDCl₃(chloroform-d); CDT (1,1′-carbonyldiimidazole); CuI (copper iodide);CuSO₄ (copper sulfate); DBU (1,8-diazabicyclo[5.4.0]undec-7-ene); DCE(dichloroethane); DCM (dichloromethane); DEAD (diethylazodicarboxylate); DMA (N,N-dimethylacetamide); DMF(N,N-dimethylformamide); DMSO (dimethyl sulfoxide); DPPA(diphenylphosphoryl azide); DTT (dithiothreitol); EDTA(ethylene-diamine-tetra-acetic acid); EGTA (ethylene-glycol-tetra-aceticacid); EtOAc (ethyl acetate); EtOH (ethanol); Hex (hexane); HOAc (aceticacid); HPLC (high-performance liquid chromatography); HRMS (highresolution mass spectrum); iPr (isopropyl); IPA (isopropyl alcohol);KOAc (potassium acetate); LCMS (liquid chromatograph-mass spectrometer);LHMDS (lithium bis(trimethylsilyl)amide); LRMS (low resolution massspectrum); MeOH (methanol); MP-B(CN)H₃ (Macroporous cyanoborohydride);NaHCO₃ (sodium bicarbonate); Na₂SO₄ (sodium sulfate); Na(OAc)₃BH (sodiumtriacetoxyborohydride); NH₄OAc (ammonium acetate); NBS(N-bromosuccinimide); NMR (nuclear magnetic resonance); PBS (phosphatebuffered saline); PCR (polymerase chain reaction); Pd(dppf)([1,1′-bis(diphenylphosphino)ferrocene]palladium); Pd(Ph₃)₄(palladium(0) tetrakis-triphenylphosphine); POCl₃ (phosphorousoxychloride); PS-DMA (polystyrene diisopropylethylamine); PS-PPh₃(polystyrene-triphenyl phosphine); TBAB (tetrabutylammonium bromide);TBAF (tetrabutylammonium fluoride); TEA (triethylamine); THF(tetrahydrofuran); TFA (trifluoroacteic acid); TFAA (trifluoroaceticanhydride); TMSCH₂N₂ (trimethylsilyldiazomethane) and Ac (acetyl); BOC(t-butoxycarbonyl); Bu (butyl); Cal (calculated); Calc'd (calculated);DIEA (diisopropylethylamine); DMAP (4-dimethylaminopyridine); EDC(N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride); Eq(equivalents); Et (ethyl); HOBt (hydroxybenzotriazole); IPA(isopropanol); LC/MS (liquid chromatograph-mass spectrometer); Me(methyl); MeCN (acetonitrile); MS (mass spectrum); NMP(N-methylpyrrolidinone); Pr (propyl); Pyr (pyridine); Sat (saturated),Tosic (p-toluenesulfonic acid) and Bn (benzyl); t-Bu (tert-butyl); dba(dibenzylideneacetone); DIPEA (diisopropylethylamine); IPAC (isopropylacetate); MTBE (tert-butyl methyl ether); OAc (acetate); RT (roomtemperature); Tf (trifluoromethanesulfonyl); Wt (weight); and XRPD(x-ray powder diffraction).

The compounds of this invention may be prepared by employing reactionsas shown in the following Reaction Schemes, in addition to otherstandard manipulations that are known in the literature or exemplifiedin the experimental procedures. The illustrative Reaction Schemes below,therefore, are not limited by the compounds listed or by any particularsubstituents employed for illustrative purposes. Substituent numberingas shown in the Reaction Schemes does not necessarily correlate to thatused in the claims and often, for clarity, a single substituent is shownattached to the compound where multiple substituents are allowed underthe definitions of Formula A hereinabove.

Synopsis of Reaction Schemes

The following Reaction Schemes, Reaction Schemes I-IV, provide usefuldetails for preparing the instant compounds. The requisite intermediatesare in some cases commercially available or can be prepared according toliterature procedures.

As illustrated in Reaction Scheme I, wherein R^(p1) and R^(p2) areindependently H or amino protective group and may be appropriatelydeprotected or replaced with other protective groups in the reactionscheme if necessary, a cycloalkyl(phenyl)acetic acid derivative is firstreacted and rearranged under Curtius-type conditions to give thecyclobutylamines I-1. Cyanation, in this case catalysed by palladium,gives nitrile I-2. Reaction of I-2 with a nucleophilic benzyl Grignardreagent and hydrolytic work-up gives ketone I-3. Condensation of I-3with aldehyde I-4 under basic conditions gives the chloronaphthyridineI-5. Displacement of the chlorine with hydrazine gives hydrazide I-6.Acylation gives acyl hydrazide I-7 which is cyclized under acidicconditions gives the triazolonaphthyridine I-8. Deprotection of I-8generates I-9.

An alternative triazolonaphthyridine synthesis is outlined in ReactionScheme II. In this case the aldehyde I-4 is first condensed with aphenylacetyl chloride under basic conditions to give thechloropyridopyrazinol II-1. The intermediate II-1 is then reacted withhydrazine and coupled with a carboxylic acid in a similar manner usedfor the conversion of I-5 to I-7 (Reaction Scheme I), to give thecarbohydrazide II-3. This is cyclized under dehydrating conditions witha reagent such as phosphorus oxychloride to give thetriazolopyridopyrazine II-4. Intermediate II-4 is then coupled with anappropriately functionalized benzene derivative, in this case boronicacid derivative, to give II-5. Deprotection of II-5 gives II-6.

Compounds of the instant invention may be prepared according to theprocedures outlined in Reaction Scheme III. In this case, diamines III-1are reacted with a methyl oxo(phenyl)acetate under basic conditions togive III-2. The intermediate III-2 may then be carried through a similarsequence of reactions as described for the synthesis of II-6 from II-1(Reaction Scheme II) to give triazolopyridopyrazine III-7.

The compound IV-1 was synthesized from the intermediate I-8 by Dimrothrearrangement and deprotection.

EXAMPLES

Examples and schemes provided are intended to assist in a furtherunderstanding of the invention. Particular materials employed, speciesand conditions are intended to be further illustrative of the inventionand do not limit the reasonable scope thereof.

Ethyl 2-(4-bromophenyl)-4-(chloromethyl)pent-4-enoate (1-1)

To a solution of ethyl (4-bromophenyl)acetate (143 g, 588 mmol) in THF(800 mL) was added LHMDS (1.13 eq in THF) at −78° C. After 30 minutes,the reaction mixture was added to a solution of3-chloro-2-chloromethyl-1-propene (147 g, 1180 mmol) in THF (500 mL) at−78° C. via cannula. The reaction was allowed to slowly warm from −78°C. to rt over 15 hours. The reaction mixture was poured into sodiumbicarbonate, extracted with EtOAc, dried over sodium sulfate, filteredand concentrated. The crude residue was purified by columnchromatography eluting with 1-20% EtOAc/Hexane. The appropriatefractions were combined and the solvent removed in vacuo to give ethyl2-(4-bromophenyl)-4-(chloromethyl)pent-4-enoate (1-1) as a clear oil. MS(M+H)⁺: 332.

Ethyl 2-(4-bromophenyl)-5-chloro-4-oxopentanoate (1-2)

Through a solution of ethyl2-(4-bromophenyl)-4-(chloromethyl)pent-4-enoate

(1-1) (7.3 g, 25 mmol) in methanol (40 mL) and CH₂Cl₂ (40 mL) at −78° C.was bubbled O₃ until the reaction turned slightly blue (6 hours). Thereaction was allowed to stir for an additional 1 hour, at which time N₂gas was bubbled through the reaction mixture until the solution wascolorless. Excess methyl sulfide (3.75 g, 60.3 mmol) was added to thereaction and the mixture was allowed to warm from −78° C. to rt. Thereaction mixture was poured into saturated sodium bicarbonate, extractedwith DCM, dried over sodium sulfate filtered and concentrated. The cruderesidue was purified by column chromatography eluting with 1-20%EtOAc/Hexane. The appropriate fractions were combined and the solventremoved in vacuo to give ethyl2-(4-bromophenyl)-5-chloro-4-oxopentanoate (1-2) as a solid. MS (M+H)⁺:153.

Ethyl 2-(4-bromophenyl)-3-[2-(chloromethyl)-1,3-dioxolan-2-yl]propanoate(1-3)

To a solution of ethyl 2-(4-bromophenyl)-5-chloro-4-oxopentanoate (1-2)(35 g, 105 mmol) and ethylene glycol (19.5 g, 315 mmol) in toluene (300mL) was added para-toluenesulfonic acid (100 mg) and the reaction washeated to reflux with a dean stark trap for 6 hours. The reactionmixture was concentrated was purified by column chromatography elutingwith 0-50% EtOAc/Hexane. The appropriate fractions were combined,concentrated, and the resulting solid was recrystallized from EtOAc andhexane to give ethyl2-(4-bromophenyl)-3-[2-(chloromethyl)-1,3-dioxolan-2-yl]propanoate (1-3)as a white solid MS (M+H)⁺: 378.

2-(4-Bromophenyl)-5,8-dioxaspiro[3.4]octane-2-carboxylic acid (1-4)

To a solution of ethyl2-(4-bromophenyl)-3-[2-(chloromethyl)-1,3-dioxolan-2-yl]propanoate (1-3)(27 g, 71.5 mmol) cooled to −78° C. in DMF (200 mL) was added NaH (8.58g, 214 mmol) and the reaction was allowed to slowly warm from −78° C. tort. Once at rt, 1N NaOH (100 mL) was added and the reaction mixture wasstirred over night. The crude reaction mixture was poured into saturatedsodium bicarbonate and washed with chloroform. The aqueous layer wasacidified with HCl, extracted with chloroform, dried over sodium sulfatefiltered and concentrated. The crude residue was purified by columnchromatography eluting with 1-50% EtOAc/Hexane. The appropriatefractions were concentrated and recrystallized from EtOAc/hexane to give2-(4-bromophenyl)-5,8-dioxaspiro[3.4]octane-2-carboxylic acid (1-4) as awhite solid. MS (M+H)⁺: 314.

tert-Butyl [2-(4-bromophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]carbamate(1-5)

To a solution of2-(4-bromophenyl)-5,8-dioxaspiro[3.4]octane-2-carboxylic acid

(1-4) (40.7 g, 130 mmol) and DIPEA (22.7 mL, 130 mmol) in tert-butanol(231 mL, 3.25 mol) was added DPPA (35.8 g, 130 mmol) and the reactionwas heated to 100° C. overnight under N₂. The reaction mixture waspoured into saturated sodium bicarbonate, extracted with EtOAc, driedover sodium sulfate, filtered and concentrated. The crude residue waspurified by column chromatography eluting with 7-50% EtOAc/Hexane. Theappropriate fractions were combined and the solvent removed in vacuo togive tert-butyl [2-(4-bromophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]carbamate(1-5). MS (M+H)⁺: 385.

tert-butyl [2-(4-cyanophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]carbamate(1-6)

To a solution of tert-butyl[2-(4-bromophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]carbamate (1-5) (21.3 g,55.5 mmol) in dioxane (100 mL) and DMF (100 mL) was added zinc cyanide(6.52 g, 55.5 mmol) and bis(tri-t-butylphosphine)palladium(0) (2.84 g,5.55 mmol) and the reaction was heated to 120° C. under N₂ for 1 hour.The reaction mixture was cooled to rt, filtered, and concentrated. Thecrude residue was purified by column chromatography eluting with 1-60%EtOAc/Hexane. The appropriate fractions were combined and the solventremoved in vacuo to give tert-butyl[2-(4-cyanophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]carbamate (1-6). MS(M+H)⁺: 331.

tert-butyl{2-[4-(phenylacetyl)phenyl]-5,8-dioxaspiro[3.4]oct-2-yl}carbamate (1-7)

To a solution of tert-butyl[2-(4-cyanophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]carbamate (1-6) (15.0 g,45.4 mmol) in THF (150 mL) at −78° C. was added isopropylmagnesiumchloride (22.7 mL, 45.4 mmol, 2M in THF). After 1 hour, benzylmagnesiumchloride (68 mL, 135 mmol, 2M in THF) was added and the reaction wasallowed to slowly warm to rt over 5 hours. The reaction mixture waspoured into saturated ammonium chloride, extracted with EtOAc, driedover sodium sulfate, filtered and concentrated. The crude residue waspurified by column chromatography eluting with 1-60% EtOAc/Hexane. Theappropriate fractions were combined and the solvent removed in vacuo togive tert-butyl{2-[4-(phenylacetyl)phenyl]-5,8-dioxaspiro[3.4]oct-2-yl}carbamate (1-7).MS (M+H)⁺: 424.

tert-butyl{2-[4-(5-chloro-3-phenyl-1,6-naphthyridin-2-yl)phenyl]-5,8-dioxaspiro[3.4]oct-2-yl}carbamate(1-9)

To a solution of tert-butyl{2-[4-(phenylacetyl)phenyl]-5,8-dioxaspiro[3.4]oct-2-yl}carbamate (1-7)(8.8 g, 20.8 mmol) in DMF (100 mL) was added potassium carbonate (14.4g, 104 mmol) and tert-butyl (2-chloro-3-formyl-4-pyridinyl)carbamate(1-8) (5.33 g, 20.8 mmol) and the reaction mixture was heated 80° C.over night. The reaction mixture was poured into saturated sodiumbicarbonate, extracted with EtOAc, dried over sodium sulfate, filteredand concentrated. The crude residue was purified by columnchromatography eluting with 1-80% EtOAc/Hexane. The appropriatefractions were combined and the solvent removed in vacuo to givetert-butyl{2-[4-(5-chloro-3-phenyl-1,6-naphthyridin-2-yl)phenyl]-5,8-dioxaspiro[3.4]oct-2-yl}carbamate(1-9). MS (M+H)⁺: 545.

tert-butyl{2-[4-(5-hydrazino-3-phenyl-1,6-naphthyridin-2-yl)phenyl]-5,8-dioxaspiro[3.4]oct-2-yl}carbamate(1-10)

To a solution of tert-butyl{2-[4-(5-chloro-3-phenyl-1,6-naphthyridin-2-yl)phenyl]-5,8-dioxaspiro[3.4]oct-2-yl}carbamate(1-9) (5 g, 9.2 mmol) in dioxane (100 mL) was added hydrazine (3.0 g, 92mmol) and the reaction mixture was heated to 100° C. for 1 hour. Thereaction mixture was poured into saturated sodium bicarbonate, extractedwith chloroform, dried over sodium sulfate, filtered and concentrated togive tert-butyl{2-[4-(5-hydrazino-3-phenyl-1,6-naphthyridin-2-yl)phenyl]-5,8-dioxaspiro[3.4]oct-2-yl}carbamate(1-10) as a yellow solid. MS (M+H)⁺: 540.

tert-butyl(2-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]oct-2-yl)carbamate(1-11)

To a solution of 1-methyl-1H-imidazole-4-carboxylic acid (7.0 g, 56mmol) in DMA (200 mL) was added EDC (11 g, 56 mmol) and HOBt (8.5 g, 56mmol) and the reaction mixture was stirred at room temperature for 1hour. Then tert-butyl{2-[4-(5-hydrazino-3-phenyl-1,6-naphthyridin-2-yl)phenyl]-5,8-dioxaspiro[3.4]oct-2-yl}carbamate(1-10) (30 g, 56 mmol) was added and the reaction stirred at roomtemperature for an additional 1 hour at which time acetic acid (17 g,280 mmol) was added and the reaction was stirred overnight at 80° C.,The reaction mixture was cooled to room temperature, quenched with 1NNaOH, poured into saturated sodium bicarbonate, extracted with EtOAc,dried (Na₂SO₄), filtered and concentrated. The crude residue waspurified by column chromatography eluting with 10% methanol in DCM. Theappropriate fractions were concentrated and the resulting solid wasrecrystallized from methanol to give tert-butyl(2-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]oct-2-yl)carbamate(1-11) as a white solid. MS (M+H)⁺: 630.

2-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]octan-2-amine(1-12)

To a solution of tert-butyl(2-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]oct-2-yl)(1-11) in DCM (50 mL) was added TFA (50 mL) at rt under N₂ for 2 hours.The reaction was quenched with 1N NaOH (650 mL), poured into saturatedsodium bicarbonate, extracted with chloroform, dried over sodiumsulfate, filtered and concentrated. The crude residue was recrystallizedfrom methanol to give2-{4-[3-(1-methyl-1/1-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]octan-2-amine(1-12) as a white solid. HRMS (M+H)⁺: observed=530.2315,calculated=530.2299.

2-(1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-3-oxocyclobutyl)-1H-isoindole-1,3(2H)-dione(1-13)

A mixture of2-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]octan-2-amine(142) (1.3 g, 2.49 mmol) and ethyl1,3-dioxo-1,3-dihydro-2H-isoindole-2-carboxylate (0.37 g, 2.49 mmol) inCHCl₃ (5 mL) was stirred at 70° C. for overnight. The mixture and DMF (5mL) and DIPEA (0.44 mL, 2.49 mmol) were heated under microwaveirradiation at 150° C. for 1 hour. The solvent was removed under reducedpressure, and to the residue was added MeOH. The resulting solid wascollected by filtration. The solid (100 mg, 0.152 mmol) and 6N HCl inMeOH (1 mL) was stirred at room temperature for 1 hour. The mixture wasquenched by aq. NaHCO₃, extracted with CHCl₃, dried (MgSO₄), filtered,and concentrated under reduced pressure to yield(2-(1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-3-oxocyclobutyl)-1H-isoindole-1,3(2H)-dione(1-13). MS (M+H)⁺: observed 616, calculated=616.

cis-3-amino-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(1-14) andtrans-3-amino-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(1-15)

A solution of(2-(1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-3-oxocyclobutyl)-1H-isoindole-1,3(2H)-dione(1-3) (203 mg, 0.330 mmol) in CH₂Cl₂ (15 mL) was added sodiumborohydride (13 mg, 0.330 mmol), and stirred at room temperature for 1hour. The mixture was quenched by aq. NaHCO₃, extracted with CHCl₃,dried (MgSO₄), filtered, and concentrated under reduced pressure to givedesired alcohol derivatives. To the micro reactor vessel was added thealcohol (204 mg, 0.330 mmol), hydrazine hydrate (170 mg, 3.4 mmol), andEtOH (2 mL), and the mixture was heated under microwave irradiation at100° C. for 10 minutes. The mixture was diluted with CHCl₃, washed withaq. NaHCO₃, dried (MgSO₄), filtered, and concentrated under reducedpressure. The resulting residue was purified by reverse phase columnchromatography (Sunfire C18) eluting with 5 to 95% acetonitrile/(0.1%TFA/water) gradient. The appropriate fractions were free based bysuspending in ethyl acetate, washed with a saturated solution of sodiumbicarbonate, followed by water, brine, dried over sodium sulfate,filtered, and concentrated in vacuo to givecis-3-amino-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(1-14), MS (M+H)⁺: observed 488, calculated=488; andtrans-3-amino-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(145), MS (M+H)⁺: observed=488, calculated=488.

cis-3-amino-1-methyl-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(1-16) andtrans-3-amino-1-methyl-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(1-17)

To a solution of(2-(1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-3-oxocyclobutyl)-1H-isoindole-1,3(2H)-dione(1-13) (100 mg, 0.162 mmol) in CH₂Cl₂ (5 mL) was added MeMgBr (2M inTHF, 0.49 mL, 0.49 mmol). After stirring for 1 h, the mixture wasquenched sat. aq. NH₄Cl, extracted with CHCl₃, dried (MgSO₄), filtered,and concentrated under reduced pressure to give a desired compound. Tothe micro reactor vessel was added the compound (103 mg, 0.162 mmol),hydrazine hydrate (80 mg, 1.6 mmol), and EtOH (2 mL), and the mixturewas heated under microwave irradiation at 100° C. for 10 minutes. Themixture was diluted with CHCl₃, washed with aq. NaHCO₃, dried (MgSO₄),filtered, and concentrated under reduced pressure. The resulting residuewas purified by reverse phase column chromatography (Sunfire C18)eluting with 5 to 95% acetonitrile/(0.1% TFA/water) gradient. Theappropriate fractions were free based by suspending in ethyl acetate,washed with a saturated solution of sodium bicarbonate, followed bywater, brine, dried over sodium sulfate, filtered, and concentrated invacuo to givecis-3-amino-1-methyl-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(1-16), MS (M+H)⁺: observed=502, calculated=502; andtrans-3-amino-1-methyl-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(1-17), MS (M+H)⁺: observed=502, calculated=502.

3,3-dimethoxy-1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine(1-18)

To a micro reactor vessel was added(2-(1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-3-oxocyclobutyl)-1H-isoindole-1,3(2H)-dione(1-13) (100 mg, 0.162 mmol), TFA (1 mL), and MeOH (5 mL), and themixture was heated under microwave irradiation at 100° C. for 10minutes. Then hydrazine hydrate (80 mg, 1.6 mmol) was added, and themixture was heated under microwave irradiation at 100° C. for 10minutes. The solvent was concentrated under reduced pressure, and theresidue was purified by reverse phase column chromatography (SunfireC18) eluting with 5 to 95% acetonitrile/(0.1% TFA/water) gradient. Theappropriate fractions were free based by suspending in ethyl acetate,washed with a saturated solution of sodium bicarbonate, followed bywater, brine, dried over sodium sulfate, filtered, and concentrated invacuo to give3,3-dimethoxy-1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine(1-18). HRMS (M+H)⁺: observed=532.2461, calculated=532.2466.

tert-butyl 5-chloro-2-oxo-3-phenyl-1,6-naphthyridine-1(2H)-carbamate(2-2)

To a solution of tert-butyl (2-chloro-3-formyl-4-pyridinyl)carbamate (10g, 39 mmol) and DBU (11.7 mL, 78 mmol) in THF (130 mL) was addedphenylacetyl chloride (5.7 mL, 43 mmol) at 0° C. The reaction wasallowed to slowly warm to room temperature for overnight. The solventwas removed under reduced pressure, and the residue was diluted withEtOAc, washed with 1N HCl, dried (MgSO₄), filtered, and concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel to give tert-butyl5-chloro-2-oxo-3-phenyl-1,6-naphthyridine-1(2H)-carbamate (2-2) as acolorless solid.

5-hydrazino-3-phenyl-1,6-naphthyridine-2(1H)-one (2-3)

A mixture of tert-butyl5-chloro-2-oxo-3-phenyl-1,6-naphthyridine-1(2H)-carbamate (2-2) (2.0 g,5.6 mmol) and hydrazine hydrate (7.2 g, 112 mmol) in 1,4-dioxane (28 mL)was heated under microwave irradiation at 120° C. for 20 minutes. Thesolvent was removed under reduced pressure, and water was added to theresidue. The resulting solid was collected by filtration to give5-hydrazino-3-phenyl-1,6-naphthyridine-2(1H)-one (2-3) as a colorlesssolid.

9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8(7H)-one(2-4)

To a mixture of 5-hydrazino-3-phenyl-1,6-naphthyridine-2(1H)-one (2-3)(1.4 g, 5.55 mmol), 2-pyrimidinecarboxylic acid (0.69 g, 5.55 mmol),HOBt (0.85 g, 5.55 mmol), and DIPEA (0.97 mL, 5.55 mmol) in DMF (50 mL)was added EDC (1.23 g, 6.66 mmol). After stirring for overnight, AcOH (5mL) was added, and the mixture was heated under microwave irradiation at150° C. for 15 minutes. The solvent was removed under reduced pressure,and the residue was diluted with MeOH, and added water. The resultingsolid was collected by filtration to give9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8(7H)-one(2-4) as a colorless solid.

8-chloro-9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridine(2-5)

To a solution of9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8(7H)-one(2-4) (1.4 g, 4.1 mmol) and DMF (0.032 mL) in acetonitrile (20 mL) wasadded POCl₃ (2.3 mL, 24.6 mmol). After stirring at 120° C. forovernight, the solvent was removed under reduced pressure, and theresidue was diluted with acetonitrile, and added water. The resultingsolid was collected by filtration to give8-chloro-9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridine(2-5) as a colorless solid.

2-(trans-3-hydroxy-3-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-1H-isoindole-1,3(2H)dione(2-6)

A mixture of8-chloro-9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridine(2-5) (37 mg, 0.103 mmol) and2-{trans-3-hydroxy-3-methyl-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]cyclobutyl}-1H-isoindole-1,3(2H)-dione(47 mg, 0.108 mmol), Na₂CO₃ (22 mg, 0.206 mmol), palladiumtetrakis(triphenylphosphine) (12 mg, 0.03 mmol) in 1,4-dioxane (3.5 mL)and water (1.2 mL) was heated under microwave irradiation at 100° C. for15 minutes. The mixture was diluted with EtOAc, washed with water, andbrine, dried (MgSO₄), filtered, and concentrated under reduced pressure.The residue was purified by column chromatography on silica gel to give2-(trans-3-hydroxy-3-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-1H-isoindole-1,3(2H)dione(2-6) as a colorless solid.

trans-3-amino-1-methyl-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(2-7)

A mixture of2-(trans-3-hydroxy-3-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-1H-isoindole-1,3(2H)dione(2-6) (25 mg, 0.040 mmol) and hydrazine hydrate (10 mg, 0.20 mmol) inEtOH (2 mL) was stirred at 100° C. for 4 hours. The solvent wasconcentrated under reduced pressure, and the residue was purified byreverse phase column chromatography (Sunfire C18) eluting with 5 to 95%acetonitrile/(0.1% TFA/water) gradient. The appropriate fractions werefree based by suspending in ethyl acetate, washed with a saturatedsolution of sodium bicarbonate, followed by water, brine, dried oversodium sulfate, filtered, and concentrated in vacuo to givetrans-3-amino-1-methyl-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(2-7) as a pale yellow solid. HRMS (M+H)⁺: observed=500.2199,calculated=500.2188.

The following compounds were prepared in a similar fashion to Example2-7, but using the appropriate starting materials:

-   trans-3-fluoro-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (2-8) HRMS (M+H)⁺: observed=488.1999, calculated=488.1992;-   cis-3-fluoro-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (2-9) MS (M+H)⁺: observed=488, calculated 488;-   3,3-difluoro-1-{-4-[9-phenyl-3-(2-primidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (2-10) HRMS (M+H)⁺: observed=506.1905, calculated=506.1905;-   trans-3-amino-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-11) HRMS (M+H)⁺: observed=486.2042, calculated=486.2034;-   cis-3-amino-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-12) HRMS (M+H)⁺: observed=486.2042, calculated=486.2039;-   trans-3-methoxy-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (2-13) HRMS (M+H)⁺: observed=500.2199, calculated=500.2189;-   trans-3-amino-1-cyclopropyl-3-{-4-[9-phenyl-3-(2-pyrimidinyl)-[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-14) HRMS (M+H)⁺: observed=526.2355, calculated=526.2341;-   cis-3-amino-1-cyclopropyl-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol (245)    MS (M+H)⁺: observed=526, calculated=526;-   3-fluoro-3-methyl-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine (246)    HRMS (M+H)⁺: observed=502.2155, calculated=502.2147;-   trans-3-amino-1-ethyl-3-{-4-[9-phenyl-3-(pyrimidin-2-yl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-17) MS (M+H)⁺: observed=550, calculated=550;-   trans-3-amino-1-methyl-3-{4-[3-(pyrimidin-2-yl)-9-(thiophen-2-yl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol    (2-18) MS (M+H)⁺: observed=506, calculated=506;-   trans-3-amino-1-methyl-3-{4-[9-phenyl-3-(pyrimidin-2-yl)[1,2,4]triazolo[4′,3′:1,2]pyrido[3,4-b]pyrazin-8-yl]phenyl}cyclobutanol    (2-19) HRMS (M+H)⁺: observed=5012139, calculated=501.2151; and-   8-{4-(trans-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl}-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-ol    (2-21) MS (M+H)⁺: observed=438, calculated=438.

6-chloro-3-phenylpyrido[2,3-b]pyrazin-2-ol (3-2)

A mixture of 6-chloro-2,3-pyrazinediamine (3-1) (15.3 g, 107 mmol),DIPEA (37.2 mL, 213 mmol), and methyl oxo(phenyl)acetate (26.2 g, 160mmol) in THF (300 mL) was stirred at room temperature for 2 days. Thesolvent was concentrated under reduced pressure, and EtOAc was added.The resulting solid was collected by filtration to give6-chloro-3-phenylpyrido[2,3-b]pyrazin-2-ol (3-2) as yellow powder.

6-hydrazino-3-phenylpyrido[2,3-b]pyrazin-2-ol (3-3)

6-chloro-3-phenylpyrido[2,3-b]pyrazin-2-ol (3-2) (26.2 g, 102 mmol) andhydrazine hydrate (26.2 g, 102 mmol) in 1,4-dioxane (400 mL) was stirredat 100° C. for 3 days. To the mixture was added EtOAc and water. Theresulting solid was collected by filtration to give6-hydrazino-3-phenylpyrido[2,3-b]pyrazin-2-ol (3-3) as yellow powder.

N-(2-hydroxy-3-phenylpyrido[2,3-b]pyrazin-6-yl)nicotinohydrazide (3-4)

To a mixture of 6-hydrazino-3-phenylpyrido[2,3-b]pyrazin-2-ol (3-3) (3.7g, 14.5 mmol) and nicotinic acid (2.1 g, 17.4 mmol) in NMP (50 mL) wasadded EDC (3.1 g, 16.0 mmol), and stirred at room temperature forovernight. To the mixture was added water, and the resulting solid wascollected by filtration to yieldN-(2-hydroxy-3-phenylpyrido[2,3-b]pyrazin-6-yl)nicotinohydrazide (3-4).

3-chloro-2-phenyl-9-(3-pyridinyl)[1,2,4]triazolo[4′,3′:1,6]pyrido-[2,3-b]pyrazine(3-5)

To a mixture ofN′-(2-hydroxy-3-phenylpyrido[2,3-b]pyrazin-6-yl)nicotinohydrazide (3-4)(5.8 g, 16.2 mmol) in acetonitrile (1.3 mL) was added POCl₃ (7.5 mL, 81mmol), and stirred at 100° C. for overnight. The solvent was removedunder reduced pressure, and to the residue was added CHCl₃ and water.The organic layer was separated, dried (Na₂SO₄), filtered, andevaporated in vacuo. To the residue was added CHCl₃, and the resultingsolid was collected by filtration to yield3-chloro-2-phenyl-9-(3-pyridinyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine(3-5) as an orange solid.

trans-3-amino-1-methyl-3-{4-[2-phenyl-9-(3-pyridinyl)[1,2,4]triazolo-[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol(3-7)

This compound was prepared in a similar manner to the procedure todescribe in scheme 4, but using the appropriate starting materials. HRMS(M+H)⁺: observed=500.2199, calculated=500.2203.

The following compounds were prepared in a similar fashion to Example3-7, but using the appropriate starting materials:

-   trans-3-amino-1-cyclopropyl-3-{4-[9-(3,4-difluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-8) HRMS (M+H)⁺: observed=561.2211, calculated=561.2214;-   trans-3-amino-1-cyclopropyl-3-[4-(2,9-diphenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]cyclobutanol    (3-9) MS (M+H)⁺: observed=525, calculated=525;

trans-3-amino-3-{4-[9-(4-chlorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol(3-10) HRMS (M+H)⁺: observed=559.2007, calculated=559.2013;

-   3-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol    (3-11) HRMS (M+H)⁺: observed=541.2331, calculated=541.2352;-   trans-3-amino-1-cyclopropyl-3-{4-[9-(4-methylphenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-12) HRMS (M+H)⁺: observed=539.2558, calculated=539.2559;-   trans-3-amino-1-methyl-3-{-4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′:3′;    1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol (3-13) MS (M+H)⁺:    observed=519, calculated=519;-   4-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol    (3-14) HRMS (M+H)⁺: observed=541.2335, calculated=541.2352;-   trans-3-amino-1-cyclopropyl-3-{4-[9-(4-fluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-15) HRMS (M+H)⁺: observed=543.2291, calculated=543.2309;-   trans-3-amino-1-cyclopropyl-3-{4-[9-(3-fluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:    1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol (3-16) HRMS    (M+H)⁺: observed=543.2305, calculated=543.2309;-   3-amino-3-{4-[9-(2-aminopyrimidin-5-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol    (3-17) HRMS (M+H)⁺: observed=542.2421, calculated=542.2417;-   3-{3-[4-(trans-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol    (3-18) HRMS (M+H)⁺: observed=515.2189, calculated=515.2195;-   trans-3-amino-3-{4-[9-(6-aminopyridin-3-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropyl    cyclobutanol (3-19) HRMS (M+H)⁺: observed=541.2462,    calculated=541.2464;-   trans-3-amino-1-cyclopropyl-3-{-4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-20) MS (M±H)⁺: observed=545, calculated=545;-   trans-3-amino-1-cyclopropyl-3-{-4-[2-phenyl-9-(pyridin-3-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-21) MS (M+-H)⁺: observed=526, calculated=526;-   trans-3-amino-1-ethyl-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-22) MS (M+H)⁺: observed=533, calculated=533;-   trans-3-amino-1-cyclopropyl-3-{4-[9-(morpholin-4-yl)-2-phenyl[1,2,4]-triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-23) MS (M+H)⁺: observed=534, calculated=534;-   trans-3-amino-3-[4-(9-ethyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-methylcyclobutanol    (3-24) MS (M+H)⁺: observed=451, calculated=451;-   trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(pyrimidin-5-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-25) MS (M+H)⁺: observed=527, calculated=527;-   trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(piperidin-1-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-27) MS (M+H)⁺: observed=532, calculated=532;-   trans-3-amino-3-{4-[9-(2-aminopyridin-4-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-methylcyclobutanol    (3-28) HRMS (M+H)⁺: observed=515.2321, calculated=515.2308;-   trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(pyrazin-2-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-29) MS (M+H)⁺: observed=527, calculated=527;-   (3-amino-3-{-4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutyl)methanol    (3-30) MS (M+H)⁺: observed=519, calculated=519;-   trans-3-amino-1-cyclopropyl-3-{4-[9-(difluoromethyl)-2-phenyl[1,2,4]triazol    o[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol (3-31) MS    (M+H)⁺: observed=499, calculated=499;-   trans-3-amino-3-{4-[9-(difluoromethyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-methylcyclobutanol    (3-32) MS (M+H)⁺: observed=473, calculated=473;-   trans-3-amino-3-{4-[9-(3-chlorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol    (3-33) HRMS (M+H)⁺: observed=559.2004, calculated=559.2013;-   trans-3-amino-1-cyclopropyl-3-{4-[9-(2-fluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-34) MS (M+H)⁺: observed=543, calculated=543;-   cis-3-amino-1-methyl-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-35) HRMS (M+H)⁺: observed=519.2111, calculated=519.2120;-   trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(trifluoromethyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-36) MS (M+H)⁺: observed=517, calculated=517;-   trans-3-amino-1-cyclopropyl-3-{4-[9-(3-methylphenyl)-2-phenyl][1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol    (3-37) HRMS (M+H)⁺: observed=539.2568, calculated=539.2559;-   cis-3-amino-1-methyl-3-{4-[9-(morpholin-4-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-38) HRMS (M+H)⁺: observed=508.2455, calculated=508.2461;-   (3-amino-3-{-4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutyl)methanol    (3-39) MS (M+H)⁺: observed=519, calculated=519;-   3-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl}benzonitrile    (3-40) HRMS (M+H)⁺: observed=550.2348, calculated=550.2355;-   cis-3-amino-1-methyl-3-{-4-[2-phenyl-9-(pyrimidin-5-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-41) HRMS (M+H)⁺: observed=501.2148, calculated=501.2151;-   2-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol    (3-42) HRMS (M+H)⁺: observed=541.2329, calculated=541.2352;-   trans-3-amino-3-[4-(9-tert-butyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol    (3-44) MS (M+H)⁺: observed=505, calculated=505;-   trans-3-amino-1-cyclopropyl-3-(4-{2-phenyl-9-[3-(trifluoromethyl)phenyl][1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol    (3-45) HRMS (M+H)⁺: observed=593.2274, calculated=593.2277;-   trans-3-amino-1-cyclopropyl-3-(4-{9-[4-(methylsulfonyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol    (3-46) HRMS (M+-H)⁺: observed=603.2173, calculated=603.2178;-   trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(1H-pyrrolo[2,3-b]pyridin-4-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-47) HRMS (M+H)⁺: observed=565.2468, calculated=565.2464;-   3,3-difluoro-1-{4-[2-phenyl-9-(1H-1,2,4-triazol-3-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanamine    (3-48) MS (M+H)⁺: observed=496, calculated=496;-   trans-3-amino-1-cyclopropyl-3-{4-[9-(2-hydroxypropan-2-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (3-49) MS (M+H)⁺: observed=507, calculated=507; and-   trans-3-amino-1-cyclopropyl-3-[4-(9-methyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]cyclobutanol    (3-50) MS (M+H)⁺: observed=463, calculated=463.

2,2,2-trifluoro-N-(2-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]oct-2-yl)acetamide(4-1)

To a mixture of 2-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]octan-2-amine(1-12′) (1.0 g, 1.90 mmol) and TEA (1.1 mL, 7.58 mmol) in CH₂Cl₂ (25 mL)was added TFAA (0.65 mL, 4.55 mmol), and stirred at 100° C. forovernight. The reaction was quenched by sat. NaHCO₃, extracted withEtOAc. The combined organic fractions were washed with brine, dried(Na₂SO₄), filtered, and the solvent was removed under reduced pressureto give 2,2,2-trifluoro-N-(2-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]oct-2-yl)acetamide(4-1) as a colorless oil.

2,2,2-trifluoro-N-(3-oxo-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-2)

To a mixture of2,2,2-trifluoro-N-(2-{9-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-5,8-dioxaspiro[3.4]oct-2-yl)acetamide(44) (620 mg, 0.99 mmol) in acetone (15 mL) was added 1N HCl (8 mL, 8.00mmol), and the mixture was stirred at 80° C. for 2 days. The precipitatewas collected by filtration and washed with water to give2,2,2-trifluoro-N-(3-oxo-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-2) as a colorless solid.

2,2,2-trifluoro-N-(3-hydroxy-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-3)

To a mixture of2,2,2-trifluoro-N-(3-oxo-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-2) (25 mg, 0.043 mmol) in CH₂Cl₂ (1 mL) and MeOH (1 mL) was addedNaBH₄ (2.0 mg, 0.052 mmol) at −78° C., and the mixture was stirred at−78° C. for 1 hour. The mixture was quenched with sat. NH₄Cl, extractedwith CHCl₃, washed with brine, dried (MgSO₄), filtered, and concentratedunder reduced pressure to give2,2,2-trifluoro-N-(3-hydroxy-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-3). MS (M+H)⁺: observed=581, calculated=581.

N-(3-(dimethylamino)-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide(4-4)

To a mixture of2,2,2-trifluoro-N-(3-oxo-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-2) (20 mg, 0.035 mmol) and dimethylamine (2M in THF, 0.052 mL, 0.104mmol) in MeOH (0.3 mL) was added Zn(BH₃CN)₂ (0.3M in MeOH, 0.6 mL, 0.18mmol), and the mixture was stirred at room temperature for overnight. Tothe mixture was added EtOAc, sat. NaHCO₃, and brine. The resulting solidwas collected by filtration to giveN-(3-(dimethylamino)-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide(4-4) as colorless solid. HRMS (M+H)⁺: observed=609.2338,calculated=609.2342.

N³,N³-dimethyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-1,3-cyclobutanediamine(4-5)

To a mixture ofN-(3-(dimethylamino)-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide(4-4) (10 mg, 0.016 mmol) in EtOH (1.0 mL) was added KOH (1M in water,0.1 mL, 0.10 mmol), and the mixture was heated under microwaveirradiation at 100° C. for 40 minutes. To the mixture was added EtOAc,sat. NaHCO₃, and brine. The resulting solid was collected by filtrationto giveN³,N³-dimethyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)-[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-1,3-cyclobutanediamine(4-5) as colorless solid. HRMS (M+H)⁺: observed=513.2515,calculated=513.2510.

The following compound was prepared in a similar fashion to Example 4-5,but using the appropriate starting materials:

-   N³-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-1,3-cyclobutanediamine    (4-6). HRMS (M+H)⁺: observed=499.2359, calculated=499.2352.

N-(3,3-difluoro-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide(4-7)

To a mixture of2,2,2-trifluoro-N-(3-oxo-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-2) (69 mg, 0.119 mmol) in CHCl₃ (2.5 mL) was added[bis(2-methoxymethyl)amino]sulfur trifluoride (0.066 mL, 0.357 mmol),and the mixture was heated under microwave irradiation at 130° C. for 30minutes. The solvent was removed under reduced pressure, and the residuewas purified by reverse phase column chromatography (Sunfire C18)eluting with 5 to 95% acetonitrile/(0.1% TFA/water) gradient. Theappropriate fractions were free based by suspending in ethyl acetate,washed with a saturated solution of sodium bicarbonate, followed bywater, brine, dried over sodium sulfate, filtered, and concentrated invacuo to giveN-(3,3-difluoro-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide(4-7) as colorless solid. MS (M+H)⁺: observed=602, calculated=602.

2,2,2-trifluoro-N-(3-hydroxy-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-N-methylacetamide(4-8)

A mixture of2,2,2-trifluoro-N-(3-hydroxy-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-3) (16 mg, 0.043 mmol), iodomethane (0.0034 mL, 0.055 mmol), andK₂CO₃ (7.6 mg, 0.055 mmol) in DMF (1 mL) was stirred at room temperaturefor 1 hour. The mixture was quenched with water, extracted with EtOAc,washed with brine, dried (MgSO₄), filtered, and concentrated underreduced pressure to give2,2,2-trifluoro-N-(3-hydroxy-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-N-methylacetamide(4-8).

3-(methylamino)-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(4-9)

This compound was prepared in a similar fashion to example 4-5, butusing the appropriate starting materials. MS (M+H)⁺: observed=500,calculated=500.

N-(3-cyano-3-hydroxy-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide(4-10)

A mixture of2,2,2-trifluoro-N-(3-oxo-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-2) (25 mg, 0.043 mmol), trimethylsilyl cyanide (0.012 mL, 0.086mmol), and zinc iodide (1.4 mg, 0.0043 mmol) in CH₂Cl₂ (1.0 mL) wasstirred at room temperature for 1 hour. To the mixture was added water,extracted with CHCl₃, washed with brine, dried (MgSO₄), filtered, andthe solvent was removed under reduced pressure. The residue was purifiedby preparative TLC, eluting with 10% MeOH in CHCl₃ to giveN-(3-cyano-3-hydroxy-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide(4-10) as a yellow solid. MS (M+H)⁺: observed=607, calculated=607.

2,2,2-trifluoro-N-(3-methylene-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-11)

A mixture of methyl(triphenyl)phosphonium bromide (111 mg, 0.311 mmol)and potassium tert-butoxide (35 mg, 0.311 mmol) in 1,4-dioxane (2 mL)was stirred at 30° C. for 30 minutes.2,2,2-trifluoro-N-(3-oxo-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-2) (60 mg, 0.104 mmol) in 1,4-dioxane (2 mL) was added, and themixture was stirred at room temperature for 15 hours. To the mixture wasadded NH₄Cl, extracted with EtOAc, washed with water and brine, dried(MgSO₄), filtered, and the solvent was removed under reduced pressure.The residue was purified by column chromatography eluting with 0-10%methanol in CHCl₃ to give2,2,2-trifluoro-N-(3-methylene-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-11) as a white solid.

2,2,2-trifluoro-N-(3-methyl-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-12)

A mixture of2,2,2-trifluoro-N-(3-methylene-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-11) (5 mg, 0.009 mmol) and palladium on carbon (10%, 10 mg) in THF (5mL) was stirred under H₂ atmosphere at room temperature for 30 minutes.The mixture was filtered through celite pad and the solvent was removedunder reduced pressure. The residue was purified by preparative TLCeluting with 5% EtOAc in hexane to give2,2,2-trifluoro-N-(3-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-12) as a white solid.

3-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine(4-13)

A mixture of2,2,2-trifluoro-N-(3-methyl-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-12) (3.3 mg, 0.006 mmol) and NaOH (5M in water, 0.30 mL) in EtOH (3mL) was stirred at 70° C. for 15 hours. The mixture was diluted withCHCl₃, washed with water and brine, dried (MgSO₄), filtered, and thesolvent was removed under reduced pressure. The residue was purified bypreparative TLC eluting with 10% MeOH in CHCl₃ to give3-methyl-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine(4-13) as a white form. HRMS (M+H)⁺: observed=484.2250,calculated=484.2243.

2,2,2-trifluoro-N-(3-hydroxy-3-(hydroxyethyl)-1-{4-[9-phenyl-3-(2-pyrimidinyl)-[1,2,4triazolo 3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide (4-14)

To a mixture of2,2,2-trifluoro-N-(3-methylene-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-11) (5 mg, 0.009 mmol) and N-methylmorpholine-N-oxide (3 mg, 0.026mmol) in THF (3 mL) was added osmium tetraoxide (1.1 mg, 0.004 mmol),and the mixture was stirred at room temperature for 3 days. To themixture was added Na₂S₂O₃, extracted with CHCl₃, washed with brine,dried (MgSO₄), filtered, filtered, and the solvent was removed underreduced pressure. The residue was purified by preparative TLC elutingwith 5% MeOH in CHCl₃ to give2,2,2-trifluoro-N-(3-hydroxy-3-(hydroxymethyl)-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-14) as a white solid.

3-amino1-(hydroxymethyl)-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(4-15)

To a mixture of2,2,2-trifluoro-N-(3-hydroxy-3-(hydroxymethyl)-1-{4-[9-phenyl-3-(2-primidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-14) (5 mg, 0.009 mmol) in EtOH (6 mL) was added NaOH (5M in water,0.5 mL, 0.50 mmol), and the mixture was refluxed for 3 hours. Thesolvent was removed under reduced pressure, and the residue was purifiedby preparative TLC eluting with 5% MeOH in CHCl₃ to give3-amino1-(hydroxymethyl)-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(4-15) as a white solid. HRMS (M+H)⁺: observed=516.2148,calculated=516.2147.

cis-1-(4-bromophenyl)-3-hydroxycyclobutanecarboxylic acid (5-1)

To a solution of isopropyl magnesium bromide (2M in THF, 127 mL, 255mmol) was added 4-bromophenylacetic acid (25 g, 116 mmol) in THF (29mL), then 2-(chloromethyl)oxirane (16.37 mL, 209 mmol) under ice bathcooling. After stirring the reaction mixture at room temperature for 45min, more isopropyl magnesium bromide (2M in THF, 116 mL, 233 mmol) wasadded over 30 min. The reaction mixture was slowly heated to 60° C. andstirred for 14 hours. The solution was then cooled to −20° C. andquenched slowly into 5N HCl. The organic layer was separated, washedwith H₂O and brine, dried (Na₂SO₄), filtered, and the solvent wasremoved under reduced pressure to give pale black and brown oil. Theresulting solid was collected by filtration to givecis-1-(4-bromophenyl)-3-hydroxycyclobutanecarboxylic acid (5-1) ascolorless powder.

(1S,5S)-5-(4-bromophenyl)-2-oxa-4-azabicyclo[3.1.1]heptan-3-one (5-2)

To a solution of cis-1-(4-bromophenyl)-3-hydroxycyclobutanecarboxylicacid (5-1) (5 g, 18.44 mmol) and Et₃N (2.57 mL, 18.44 mmol) in t-BuOH(92 mL) and 1,4-Dioxane (92 mL) was added diphenyl azidophosphate (3.97mL, 18.44 mmol), and stirred at 80° C. for 3 hours. The solvent wasremoved under reduced pressure. The residue was diluted with EtOAc,washed with sat NaHCO₃aq and brine, dried (Na₂SO₄), filtered, andconcentrated under reduced pressure. The solid was washed with EtOAc togive (1S,5S)-5-(4-bromophenyl)-2-oxa-4-azabicyclo[3.1.1]heptan-3-one(5-2) as colorless powder.

cis-3-amino-(4-bromophenyl)cyclobutanol (5-3)

A solution of aqueous KOH (4M, 167 mL, 668 mmol) and(1S,5S)-5-(4-bromophenyl)-2-oxa-4-azabicyclo[3.1.1]heptan-3-one (5-2) (3g, 11.2 mmol) in 2-Propanol (167 mL) was stirred at 100° C. forovernight. The solvent was removed under reduced pressure. The residuewas diluted with CHCl₃, washed with brine, dried (Na₂SO₄), filtered, andthe solvent was removed under reduced pressure to givecis-3-amino-(4-bromophenyl)cyclobutanol (5-3) as colorless oil.

tert-butyl [cis-1-(4-bromophenyl)-3-hydroxycyclobutyl]carbamate (5-4)

To a mixture of cis-3-amino-(4-bromophenyl)cyclobutanol (5-3) (2.7 g,11.2 mmol) and TEA (6.2 mL, 44.8 mmol) in THF (53 mL) was addeddi-tert-butyl dicarbonate (5.20 mL, 22.38 mmol), and the mixture wasstirred at room temperature for 2 hours. The solvent was removed underreduced pressure, and the residue was diluted with EtOAc, washed withsat NaHCO₃ aq and brine, dried (Na₂SO₄), filtered, and concentratedunder reduced pressure. The residue was purified by silica gelchromatography (0˜100% EtOAc in hexane) to give tert-butyl[cis-1-(4-bromophenyl)-3-hydroxycyclobutyl]carbamate (5-4) as colorlessoil.

tert-butyl [1-(4-bromophenyl)-3-oxocyclobutyl]carbamate (5-5)

To a solution of tert-butyl[cis-1-(4-bromophenyl)-3-hydroxycyclobutyl]carbamate (5-4) (3.0 g, 8.77mmol), DMSO (6.8 mL, 96 mmol) and TEA (6.1 mL, 43.8 mmol) in CHCl₃ (90mL) was added sulfur trioxide pyridine complex (5.6 g, 35.1 mmol) at 0°C., and the mixture was stirred at 0° C. for 10 minutes and at roomtemperature for overnight. The solvent was removed under reducedpressure, and the residue was diluted with water, extracted with Et₂O,washed with brine, dried (Na₂SO₄), filtered, and the solvent was removedunder reduced pressure. The residue was purified by silica gelchromatography (0˜100% EtOAc in hexane) to give tert-butyl[1-(4-bromophenyl)-3-oxocyclobutyl]carbamate (5-5) as colorless powder.

tert-butyl [1-(4-bromophenyl)-3,3-difluorocyclobutyl]carbamate (5-6)

To a solution of tert-butyl [1-(4-bromophenyl)-3-oxocyclobutyl]carbamate(5-5) (340 mg, 1.00 mmol) in CHCl₃ (10 mL) was addedbis(2-methoxyethyl)amino sulfur trifluoride (0.93 mL, 5.04 mmol), andthe mixture was stirred at room temperature for overnight. The mixturewas quenched with sat. NaHCO₃ aq., extracted with CHCl₃, washed withbrine, dried (MgSO₄), filtered, and the solvent was removed underreduced pressure. The residue was purified by silica gel chromatography(0-40% EtOAc in hexane) to give tert-butyl[1-(4-bromophenyl)-3,3-difluorocyclobutyl]carbamate (5-6) as pale yellowsolid.

3,3-difluoro-1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl])phenyl}cyclobutanamine5-12

This compound was synthesized in a manner similar to the procedure inScheme 1, but using the appropriate starting materials such as thecompound 5-7. HRMS (M+H)⁺: observed=508.2061, calculated=508.2059.

The following compounds were prepared in a similar fashion to Example5-12, but using the appropriate starting materials:

-   3,3-difluoro-1-{-4-[9-phenyl-3-(trifluoromethyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (5-13) HRMS (M+H)⁺: observed=496.1561, calculated=496.1559;-   3,3-difluoro-1-{4-[9-phenyl-3-(1H-1,2,3-triazol-4-yl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (5-14) HRMS (M+H)⁺: observed=495.1857, calculated=495.1843;-   3,3-difluoro-1-{4-[3-(1H-indazol-3-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]-phenyl}cyclobutanamine    (5-15) HRMS (M+H)⁺: observed=544.2061, calculated=544.2041;-   3,3-difluoro-1-{4-[3-(5-methyl-1H-1,2,4-triazol-3-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine    (5-16) HRMS (M+H)⁺: observed=509.2014, calculated=509.2013; and-   8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridine-3-carboxamide    (5-17) HRMS (M+H)⁺: observed=471.1745, calculated=471.1733.

tert-butyl{1-[4-(3-amino-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl)phenyl]-3,3-difluorocyclobutyl}carbamate(5-18)

A solution of{3,3-difluoro-1-[4-(5-hyderazino-3-phenyl-1,6-naphthyridin-2-yl)phenyl]cyclobutyl}carbamate(5-10) (13 mg, 0.025 mmol), cyanogen bromide (9.5 mg, 0.090 mmol), andNa₂CO₃ (13 mg, 0.12 mmol) in EtOH (0.5 mL) was stirred at roomtemperature for overnight. The solvent was removed under reducedpressure and the residue was purified by silica gel chromatography(0-10% MeOH in CHCl₃) to give tert-butyl{1-[4-(3-amino-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl)phenyl]-3,3-difluorocyclobutyl}carbamate(5-18) as pale yellow oil.

8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-amine(5-19)

To a mixture of{1-[4-(3-amino-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl)phenyl]-3,3-difluorocyclobutyl}carbamate(5-18) (4 mg, 0.007 mmol) in CHCl₃ (0.5 mL) was added TFA (0.5 mL), andthe mixture was stirred at room temperature for 1 hour. The solvent wasconcentrated under reduced pressure and the residue was purified byreverse phase column chromatography (Sunfire C18) eluting with 5 to 95%acetonitrile/(0.1% TFA/water) gradient. The appropriate fractions werefree based by suspending in ethyl acetate, washed with a saturatedsolution of sodium bicarbonate, followed by water, brine, dried oversodium sulfate, filtered, and concentrated in vacuo to give8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-amine(5-19) as a yellow oil. HRMS (M+H)⁺: observed=443.1796,calculated=443.1801.

tert-butyl{3,3-difluoro-1-[4-(3-hydroxy-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl)phenyl]cyclobutyl}carbamate(5-20)

A solution of{3,3-difluoro-1-[4-(5-hyderazino-3-phenyl-1,6-naphthyridin-2-yl)phenyl]cyclobutyl}carbamate(540) (14 mg, 0.027 mmol) and CDI (11 mg, 0.066 mmol) in 1,4-dioxane(0.55 mL) was stirred at 95° C. for overnight. The solvent was removedunder reduced pressure and the residue was purified by silica gelchromatography (0-10% MeOH in CHCl₃) to give tert-butyl{3,3-difluoro-1-[4-(3-hydroxy-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl)phenyl]cyclobutyl}carbamate(5-20) as pale yellow oil.

8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-ol(5-21)

To a mixture of tert-butyl{3,3-difluoro-1-[4-(3-hydroxy-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl)phenyl]cyclobutyl}carbamate(5-20) (11 mg, 0.021 mmol) in CHCl₃ (0.5 mL) was added TFA (0.5 mL), andthe mixture was stirred at room temperature for 1 hour. The solvent wasconcentrated under reduced pressure and the residue was purified byreverse phase column chromatography (Sunfire C18) eluting with 5 to 95%acetonitrile/(0.1% TFA/water) gradient. The appropriate fractions werefree based by suspending in ethyl acetate, washed with a saturatedsolution of sodium bicarbonate, followed by water, brine, dried oversodium sulfate, filtered, and concentrated in vacuo to give8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-ol(5-21) as a yellow oil. HRMS (M+H)⁺: observed=444.1636,calculated=444.1653.

3-amino-3-{4-[9-phenyl-2-(2-pyrimidinyl)[1,2,4]triazolo[5,1-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(6-1)

To a mixture of2,2,2-trifluoro-N-(3-hydroxy-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide(4-3) (10 mg, 0.016 mmol) in EtOH (1.0 mL) was added NaOH (2M in water,1.0 mL, 2.00 mmol), and the mixture was heated under microwaveirradiation at 100° C. for 1 hour. The mixture was diluted with water,extracted with CHCl₃, washed with brine, dried (MgSO₄), filtered, andconcentrated. The residue was purified by preparative TLC, eluting with10% MeOH in CHCl₃ to give3-amino-3-{-4-[9-phenyl-2-(2-pyrimidinyl)[1,2,4]triazolo[5,1-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol(6-1) as a colorless solid. MS (M+H)⁺: observed=486, calculated=486.

2(4-bromophenyl)-5,8-dioxaspiro[3.4]octan-2-amine (7-1)

A mixture of tert-butyl[2-(4-bromophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]carbamate (1-5) (3.7 g,9.63 mmol) in TFA (20 mL) was stirred at room temperature for 1 hour.The mixture was poured into a sat. NaHCO₃ solution, extracted withCHCl₃, dried (Na₂SO₄), filtered, and concentrated under reduced pressureto give 2-(4-bromophenyl)-5,8-dioxaspiro[3.4]octan-2-amine (7-1).

2-[4-(4-bromophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]-1H-isoindole-1,3(2H)-dione(7-2)

A mixture of 2-(4-bromophenyl)-5,8-dioxaspiro[3.4]octan-2-amine (7-1)(2.7 g, 9.50 mmol), ethyl1,3-dioxo-1,3-dihydro-2H-isoindole-2-carboxylate (4.2 g, 19.0 mmol), andTEA (5.3 mL, 38.0 mmol) in CHCl₃ (50 mL) was stirred at 70° C. forovernight. The solvent was evaporated under reduced pressure, and to theresidue was added MeOH. The resulting solid was collected by filtrationto give2-[2-(4-bromophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]-1H-isoindole-1,3(2H)-dione(7-2) as a colorless solid.

2-[1-(4-bromophenyl)-3-oxocyclobutyl]-1H-isoindole-1,3(2H)-dione (7-3)

A mixture of2-[2-(4-bromophenyl)-5,8-dioxaspiro[3.4]oct-2-yl]-1H-isoindole-1,3(2H)-dione(7-2) (3.1 g, 7.48 mmol) and p-toluenesulfonic acid monohydrate (0.14 g,0.75 mmol) in acetone (50 mL) was refluxed for 2 days. The cooledmixture was diluted with EtOAc, washed with sat. NaHCO₃, dried (MgSO₄),filtered, and concentrated under reduced pressure. The residue waspurified by silica gel chromatography, eluting with 0˜100% EtOAc inhexane, to give2-[1-(4-bromophenyl)-3-oxocyclobutyl]-1H-isoindole-1,3(2H)-dione (7-3)as a colorless form.

2-[trans-1-(4-bromophenyl)-3-hydroxy-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-4) and2-[cis-1-(4-bromophenyl)-3-hydroxy-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-5)

To a solution of2-[1-(4-bromophenyl)-3-oxocyclobutyl]-1H-isoindole-1,3(2H)-dione (7-3)(100 mg, 0.270 mmol) in CH₂Cl₂ (2 mL) was added methylmagnesium bromide(3M in THF, 0.11 mL, 0.33 mmol) at −20° C. After 1 hour stirring,aqueous citric acid was added to the mixture, extracted with CHCl₃,dried (MgSO₄), filtered, and concentrated under reduced pressure. Theresidue was purified by silica gel chromatography, eluting with 0˜100%EtOAc in hexane, to give2-[trans-1-(4-bromophenyl)-3-hydroxy-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-4) and2-[cis-1-(4-bromophenyl)-3-hydroxy-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-5).

2-{trans-3-hydroxy-3-methyl-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]cyclobutyl}-1H-isoindole-1,3(2H)-dione(7-6)

A mixture of2-[trans-1-(4-bromophenyl)-3-hydroxy-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-4) (40 mg, 0.104 mmol), bis(pinacolato)diboron (32 mg, 0.124 mmol),[1,2-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complex withCH₂Cl₂ (8.5 mg, 0.0010 mmol), and KOAc (20 mg, 0.207 mmol) in DMSO (2mL) was stirred at 80° C. for 3 hours. The mixture was diluted withEtOAc, filtered through celite pad, washed with water, dried (MgSO₄),filtered, and concentrated. The residue was purified by silica gelchromatography, eluting with 0˜100% EtOAc in hexane, to give2-{trans-3-hydroxy-3-methyl-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]cyclobutyl}-1H-isoindole-1,3(2H)-dione(7-6) as a colorless amorphous.

2-{cis-3-hydroxy-3-methyl-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]cyclobutyl}-1H-isoindole-1,3(2H)-dione(7-7)

This compound was prepared in a same manner to example 7-6, but using2-[cis-1-(4-bromophenyl)-3-hydroxy-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-5) as a starting material.

The following compounds were prepared in a similar fashion to Example7-6, but using the appropriate starting materials:

-   2-{trans-3-cyclopropyl-3-hydroxy-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]cyclobutyl}-1H-isoindole-1,3(2H)-dione    (7-8); and-   2-{cis-3-cyclopropyl-3-hydroxy-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]cyclobutyl}-1H-isoindole-1,3(2H)-dione    (7-9).

2-[trans-1-(4-bromophenyl)-3-hydroxycyclobutyl]-1H-isoindole-1,3(2H)-dione(7-10)

To a solution of2-[1-(4-bromophenyl)-3-oxocyclobutyl]-1H-isoindole-1,3(2H)-dione (7-3)(216 mg, 0.583 mmol) in THF (2 mL) was added L-selectride (1M in THF,0.59 mL, 0.59 mmol) at −78° C. After stirring 45 minutes, the reactionwas quenched by sat. aq. NH₄Cl, and the mixture was extracted withEtOAc. The combined organic fractions were dried (MgSO₄), filtered, andconcentrated under reduced pressure. The residue was purified by silicagel chromatography, eluting with 0˜100% EtOAc in hexane, to give2-[trans-1-(4-bromophenyl)-3-hydroxycyclobutyl]-1H-isoindole-1,3(2H)-dione(7-10) as a colorless solid.

2-[cis-1-(4-bromophenyl)-3-fluorocyclobutyl]-1H-isoindole-1,3(2H)-dione(7-11)

To a solution of2-[trans-1-(4-bromophenyl)-3-hydroxycyclobutyl]-1H-isoindole-1,3(2H)-dione(7-8) (162 mg, 0.435 mmol) in CHCl₃ (2 mL) was addedbis(methoxyethyl)amino sulfur trifluoride (0.09 mL, 0.522 mmol), and themixture was stirred at room temperature for 1 hour. The reaction wasquenched by sat. aq. NaHCO₃, and the mixture was extracted with EtOAc.The combined organic fractions were dried (MgSO₄), filtered, andconcentrated under reduced pressure. The residue was purified by silicagel chromatography, eluting with 0˜100% EtOAc in hexane, to give2-[cis-1-(4-bromophenyl)-3-fluorocyclobutyl]-1H-isoindole-1,3(2H)-dione(7-11) as a colorless oil.

2-{cis-3-fluoro-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxabololan-2-yl)phenyl]cyclobutyl}-1H-isoindole-1,3(2H)-dione(7-12)

This compound was prepared in a same manner to example 7-6, but using2-[cis-1-(4-bromophenyl)-3-fluorocyclobutyl]-1H-isoindole-1,3(2H)-dione(7-11) as a starting material.

tert-butyl{trans-3-fluoro-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxabololan-2-yl)phenyl]cyclobutyl}carbamate(7-14)

This compound was prepared in a same manner to example 7-10, but usingthe appropriate starting materials.

tert-butyl{3,3-difluoro-1-[4,4,5,5-tetramethyl-1,3,2-dioxabololan-2-yl)phenyl]cyclobutyl}carbamate(7-15)

This compound was prepared in a same manner to example 7-6, but usingtert-butyl [1-(4-bromophenyl)-3,3-difluorocyclobutyl]carbamate (5-6) asa starting material.

2-[trans-1-(4-bromophenyl)-3-methoxycyclobutyl}-1H-isoindole-1,3(2H)-dione(7-16)

To a solution of2-[trans-1-(4-bromophenyl)-3-hydroxycyclobutyl]-1H-isoindole-1,3(2H)-dione(7-10) (202 mg, 0.543 mmol) in DMF (2 mL) was added sodium hydride (60%,26 mg, 0.651 mmol). After stirring for 30 minutes, iodomethane (0.051mL, 0.814 mmol) was added, and the mixture was stirred at roomtemperature for 1 hour. To the mixture was added aq. ammonium chloride,and the mixture was extracted with EtOAc. The combined organic fractionswere dried (MgSO₄), filtered, and concentrated under reduced pressure.The residue was purified by silica gel column chromatography, eluting0˜100% EtOAc in hexane to give2-[trans-1-(4-bromophenyl)-3-methoxycyclobutyl}-1H-isoindole-1,3(2H)-dione(7-16) as a colorless solid.

2-{trans-3-methoxy-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxabololan-2-yl)phenyl]cyclobutyl}-1H-isoindole-1,3(2H)-dione(747)

This compound was prepared in a same manner to example 7-6, but using2-[trans-1-(4-bromophenyl)-3-methoxycyclobutyl}-1H-isoindole-1,3(2H)-dione(7-16) as a starting material.

2-[1-(4-bromophenyl)-3-fluoro-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-18)

To a solution of2-[cis-1-(4-bromophenyl)-3-hydroxy-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-5) (68 mg, 0.176 mmol) in CHCl₃ (2 mL) was addedbis(methoxyethyl)amino sulfur trifluoride (0.08 mL, 0.434 mmol), and themixture was stirred at room temperature for 4 hours. The mixture wasdiluted with CHCl₃, washed with water, dried (Na₂SO₄), filtered, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography, eluting 0-60% EtOAc in hexane to give2-[1-(4-bromophenyl)-3-fluoro-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-18) as a colorless solid.

2-{3-fluoro-3-methyl-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxabololan-2-yl)phenyl]cyclobutyl}-1H-isoindole-1,3(2H)-dione(749)

This compound was prepared in a same manner to example 7-6, but using2-[1-(4-bromophenyl)-3-fluoro-3-methylcyclobutyl]-1H-isoindole-1,3(2H)-dione(7-18) as a starting material.

N′-(2-oxo-3-phenyl-1,2-dihydropyrido[2,3-b]pyrazin-6-yl)morpholine-4-carbohydrazide(8-1)

To a mixture of 6-hydrazino-3-phenylpyrido[2,3-b]pyrazin-2-ol (3-3) (200mg, 0.79 mmol) and DIEA (0.55 mL, 3.16 mmol) in NMP (8 mL) was addedmorpholine-4-carbonyl chloride (354 mg, 2.37 mmol), and stirred at roomtemperature for 1 hour. To the mixture was added water and Et₂O, and theprecipitate was collected by filtration to yieldN′-(2-oxo-3-phenyl-1,2-dihydropyrido[2,3-b]pyrazin-6-yl)morpholine-4-carbohydrazide(8-1) as a pale yellow solid.

3,9-dichloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine(8-2)

A mixture ofN′-(2-oxo-3-phenyl-1,2-dihydropyrido[2,3-b]pyrazin-6-yl)morpholine-4-carbohydrazide(8-1) (165 mg, 0.450 mmol) and phosphorus oxychloride (5 mL) was stirredat 150° C. for 6 h. The solvent was removed under reduced pressure, andto the residue was added CHCl₃ and sat. aq. NaHCO₃. The organic layerwas separated, dried (MgSO₄), filtered, and evaporated in vacuo. Theresidue was purified by column chromatography eluting with 0-5%MeOH/CHCl₃. The appropriate fractions were combined and the solventremoved in vacuo to give3,9-dichloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine(8-2) as a pale yellow foam.

tert-butyl{trans-1-[4-(9-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-3-cyclopropyl-3-hydroxycyclobutyl}carbamate(8-3)

This compound was prepared in a similar manner to the procedure todescribe in scheme 5, but using the appropriate starting materials.

trans-3-amino-3-[4-(9-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol(8-4)

To a mixture of tert-butyl{trans-1-[4-(9-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-3-cyclopropyl-3-hydroxycyclobutyl}carbamate(8-3) (280 mg, 0.48 mmol) in EtOH (5 mL) was added 5N HCl (5 mL), andstirred at 50° C. for 20 min. The solvent was removed under reducedpressure to givetrans-3-amino-3-[4-(9-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol(8-4) as a colorless solid.

trans-3-amino-1-cyclopropyl-3-{4-[9-(dimethylamino)-2-phenyl-[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol(8-5)

A mixture oftrans-3-amino-3-[4-(9-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol(8-4) (30 mg, 0.062 mmol) and dimethylamine hydrochloride (51 mg, 0.62mmol), and triethylamine (0.087 mL, 0.62 mmol) in NMP (1.5 mL) wasstirred at 180° C. for 30 minutes. To a cooled mixture was added water,and filtered through membrane filter. The residue was purified byreverse phase column chromatography (Sunfire C18) eluting with 5 to 95%acetonitrile/(0.1% formic acid/water) gradient. The appropriatefractions were free based by suspending in ethyl acetate, washed with asaturated solution of sodium bicarbonate, followed by water, brine,dried over sodium sulfate, filtered, and concentrated under reducedpressure to givetrans-3-amino-1-cyclopropyl-3-{4-[9-(dimethylamino)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol(8-5). MS (M+H)⁺: observed=492, calculated=492.

The following compounds were prepared in a similar fashion to Example8-5, but using the appropriate starting materials:

-   trans-3-amino-1-cyclopropyl-3-{4-[9-(methylamino)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (8-6) MS (M+H)⁺: observed=478, calculated=478;-   trans-3-amino-3-[4-(9-amino-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol    (8-7) MS (M+H)⁺: observed=464, calculated=464;-   trans-3-amino-3-{4-[9-(azetidin-1-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol    (8-8) MS (M+H)⁺: observed=504, calculated=504;-   trans-3-amino-3-[4-(9-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol    (8-9) MS (M+H)⁺: observed=483, calculated=483;-   trans-3-amino-1-cyclopropyl-3-{4-[9-(methylsulfanyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol    (8-10) MS (M+H)⁺: observed=495, calculated=495;-   trans-3-amino-1-cyclopropyl-3-[4-(9-methoxy-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]cyclobutanol    (8-11) MS (M+H)⁺: observed=479, calculated=479; and-   3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-ol    (8-12) MS (M+H)⁺: observed=465, calculated=465.

3-hydroxy-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-carboxamide(9-1)

To a mixture of 6-hydrazinyl-3-phenylpyrido[2,3-b]pyrazin-2(1H)-one(3-3) (300 mg, 1.19 mmol), HOBt (218 mg, 1.42 mmol) and oxamic acid (130mg, 1.42 mmol) in NMP (12 mL) was added EDC (272 mg, 1.42 mmol) at roomtemperature and the mixture was stirred for 30 min. After addition ofTFA (0.091 mL, 1.19 mmol), the mixture was heated under microwaveirradiation at 150° C. for 30 minutes. To a mixture was added water (60mL) and the precipitate was collected by filtration to give3-hydroxy-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-carboxamide(9-1) as a yellow powder.

9-carbamoyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yltrifluoromethanesulfonate (9-2)

To a mixture of3-hydroxy-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-carboxamide(9-1) (52 mg, 0.17 mmol) and potassium carbonate (28 mg, 0.20 mmol) inDMF (1.0 mL) was added1,1,1-trifluoro-N-phenyl-N-[(trifluoromethyl)sulfonyl]methanesulfonamide(67 mg, 0.19 mmol). After stirring for 3 hr, the mixture was dilutedwith CHCl₃, washed with sat. aq. NaHCO₃ and brine, dried (MgSO₄),filtered, and concentrated under reduced pressure to give9-carbamoyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yltrifluoromethanesulfonate (9-2).

3-{4-[trans-3-cyclopropyl-1-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-3-hydroxycyclobutyl]phenyl}-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine-9-carboxamide(9-3)

A mixture of9-carbamoyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yltrifluoromethanesulfonate (9-2) (40 mg, 0.091 mmol),{4-[trans-3-cyclopropyl-1-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-3-hydroxycyclobutyl]phenyl}boronicacid (7-20) (41 mg, 0.11 mmol), cesium carbonate (60 mg, 0.183 mmol),palladium tetrakis(triphenylphosphine) (21 mg, 0.018 mmol) in1,4-dioxane (0.8 mL) was stirred at 110° C. for 1 hr. The mixture wasdiluted with CHCl₃/IPA, washed with sat. aq. NaHCO₃ and brine, dried(MgSO₄), filtered, and concentrated under reduced pressure to give3-{4-[trans-3-cyclopropyl-1-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-3-hydroxycyclobutyl]phenyl}-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine-9-carboxamide(9-3).

3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-carboxamide(9-4)

A mixture of3-{-4-[trans-3-cyclopropyl-1-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-3-hydroxycyclobutyl]phenyl}-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine-9-carboxamide(9-3) (40 mg, 0.064 mmol) and hydrazine hydrate (50 mg, 1.0 mmol) inEtOH (1 mL) was stirred at 50° C. for 30 minutes. The solvent wasevaporated in vacuo, and the residue was diluted with EtOH (1 mL) andstirred at 130° C. for 30 minutes. The solvent was removed under reducedpressure, and the residue was diluted with CHCl₃/IPA, washed with sat.aq. NaHCO₃ and brine, dried (MgSO₄), filtered, and concentrated underreduced pressure. The residue was purified by column chromatographyeluting with 0-20% MeOH/CHCl₃. The appropriate fractions were combinedand the solvent removed in vacuo to give3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′;1,6]pyrido[2,3-b]pyrazin-9-carboxamide (9-4). MS (M+H)⁺: observed=492,calculated=492.

{4-[trans-3-cyclopropyl-1-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-3-hydroxycyclobutyl]phenyl}boronicacid (7-20)

To a mixture of2-{trans-3-cyclopropyl-3-hydroxy-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]cyclobutyl-1H-isoindole-1,3(2H)-dione(7-8) (306 mg, 0.665 mmol) in acetone (44 mL) and water (22 mL) wasadded sodium periodate (860 mg, 4.01 mmol) and ammonium acetate (314 mg,4.07 mmol), and stirred at room temperature for overnight. To a mixturewas added water (100 mL) and the resulting solid was collected byfiltration to give{4-[trans-3-cyclopropyl-1-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-3-hydroxycyclobutyl]phenyl}boronicacid (7-20).

2-chloro-N′-(2-oxo-3-phenyl-1,2-dihydropyrido[2,3-b]pyrazin-6-yl)-acetohydrazide(10-1)

To a mixture of 6-hydrazinyl-3-phenylpyrido[2,3-b]pyrazin-2(1H)-one(3-3) (300 mg, 1.19 mmol) in CHCl₃ (20 mL) and pyridine (10 mL) wasadded chloroacetic anhydride (304 mg, 1.78 mmol). After stirring for 30min, the solvent was removed under reduced pressure. The residue wasdissolved in MeOH and then water was added. The precipitate wascollected by filtration to give2-chloro-N′-(2-oxo-3-phenyl-1,2-dihydropyrido[2,3-b]pyrazin-6-yl)acetohydrazide(10-1) as a colorless solid.

3-chloro-9-(chloromethyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]-pyrazine(10-2)

A mixture of2-chloro-N′-(2-oxo-3-phenyl-1,2-dihydropyrido[2,3-b]pyrazin-6-yl)acetohydrazide(104) (230 mg, 0.698 mmol) and phosphoryl chloride (4 mL, 42.9 mmol) washeated under microwave irradiation at 150° C. for 1.5 hours. The solventwas evaporated, and the residue was diluted with EtOAc, washed with sat.NaHCO₃, dried (MgSO₄), filtered, and concentrated under reduced pressureto give3-chloro-9-(chloromethyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine(10-2).

1-(3-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl)-N,N-dimethylmethanamine(10-3)

A mixture of3-chloro-9-(chloromethyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine(10-2) (840 mg, 0.254 mmol) and dimethylamine (2M in THF, 5 mL, 10 mmol)was stirred at room temperature for 2 hours. The solvent was evaporatedto give1-(3-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl)-N,N-dimethylmethanamine(10-3).

2-[trans-3-cyclopropyl-1-(4-{9-[(dimethylamino)methyl]-2-phenyl-[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)-3-hydroxycyclobutyl]-1H-isoindole-1,3(2H)-dione(10-4)

This compound was prepared in a similar manner to the procedure todescribe in scheme 5, but using the appropriate starting materials.

trans-3-amino-1-cyclopropyl-3-(4-{9-[(dimethylamino)methyl]-2-phenyl-[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol(10-5)

This compound was prepared in a similar manner to the procedure todescribe in scheme 4, but using the appropriate starting materials. MS(M+H)⁺: observed=506, calculated=506.

The following compound was prepared in a similar fashion to Example10-5, but using the appropriate starting materials:

-   N-({3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl}methyl)acetamide    (10-6) MS (M+H)⁺: observed=520, calculated=520.

methyloxo[2-oxo-3-phenyl-1,2-dihydropyrido[2,3-b]pyrazin-6-yl)hydrazinyl]acetate(11-1)

To a mixture of 6-hydrazinyl-3-phenylpyrido[2,3-b]pyrazin-2(1H)-one(3-3) (500 mg, 1.97 mmol) in DMF (10 mL) and pyridine (10 mL) was addedmethyl chloroglyoxylate (290 mg, 2.37 mmol). After stirring for 30 min,the solvent was removed under reduced pressure. The residue wasdissolved in MeOH and then water was added. The precipitate wascollected by filtration to give methyloxo[2-oxo-3-phenyl-1,2-dihydropyrido[2,3-b]pyrazin-6-yl)hydrazinyl]acetate(11-1).

methyl3-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine-9-carboxylate(11-2)

This compound was prepared in a similar fashion to Example 3-5, butusing the appropriate starting materials.

3-chloro-N-ethyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine-9-carboxamide(11-3)

A mixture of methyl3-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine-9-carboxylate(11-2) (80 mg, 0.235 mmol) and ethylamine (2M in THF, 5 mL, 10 mmol) wasstirred at room temperature for overnight. The solvent was evaporatedand the residue was purified by column chromatography eluting with 0-20%EtOH in EtOAc to give3-chloro-N-ethyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazine-9-carboxamide(11-3).

3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-N-ethyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-carboxamide(11-5)

This compound was prepared in a similar manner to the procedure todescribe in scheme 4, but using the appropriate starting materials. MS(M+H)⁺: observed=520, calculated=520.

Example 1 Cloning of the Human Akt Isoforms and Delta-PH-Akt1 (PH DomainDeleted AKT1)

The pS2neo vector (deposited in the ATCC on Apr. 3, 2001 as ATCCPTA-3253) was prepared as follows: The pRmHA3 vector (prepared asdescribed in Nucl. Acid Res. 16:1043-1061 (1988)) was cut with BglII anda 2734 bp fragment was isolated. The pUChsneo vector (prepared asdescribed in EMBO J. 4:167-171 (1985)) was also cut with BglII and a4029 bp band was isolated. These two isolated fragments were ligatedtogether to generate a vector termed pS2neo-1. This plasmid contains apolylinker between a metallothionine promoter and an alcoholdehydrogenase poly A addition site. It also has a neo resistance genedriven by a heat shock promoter. The pS2neo-1 vector was cut with Psp5IIand BsiWI. Two complementary oligonucleotides were synthesized and thenannealed (CTGCGGCCGC (SEQ. ID. NO.: 1) and GTACGCGGCCGCAG (SEQ. ID. NO.:2)). The cut pS2neo-1 and the annealed oligonucleotides were ligatedtogether to generate a second vector, pS2neo. Added in this conversionwas a NotI site to aid in the linearization prior to transfection intoS2 cells.

Human Akt1 gene was amplified by PCR (Clontech) out of a human spleencDNA (Clontech) using the 5′ primer:5′CGCGAATTCAGATCTACCATGAGCGACGTGGCTATTGTG 3′ (SEQ. ID. NO.: 3), and the3′ primer: 5′CGCTCTAGAGGATCCTCAGGCCGTGCTGCTGGC3′ (SEQ. ID. NO.: 4). The5′ primer included an EcoRI and BglII site. The 3′ primer included anXbaI and BamHI site for cloning purposes. The resultant PCR product wassubcloned into pGEM3Z (Promega) as an EcoRI/Xba I fragment. Forexpression/purification purposes, a middle T tag was added to the 5′ endof the full length Akt1 gene using the PCR primer:5′GTACGATGCTGAACGATATCTTCG 3′ (SEQ. ID. NO.: 5). The resulting PCRproduct encompassed a 5′ KpnI site and a 3′ BamHI site which were usedto subclone the fragment in frame with a biotin tag containing insectcell expression vector, pS2neo.

For the expression of a pleckstrin homology domain (PH) deleted (Δaa4-129, which includes deletion of a portion of the Akt1 hinge region)version of Akt1, PCR deletion mutagenesis was done using the full lengthAkt1 gene in the pS2neo vector as template. The PCR was carried out in 2steps using overlapping internal primers(5′GAATACATGCCGATGGAAAGCGACGGGGCTGAAGAGATGGAGGTG 3′ (SEQ. ID. NO.: 6),and 5′CCCCTCCATCTCTTCAGCCCCGTCGCTTICCATCGGCATG TATTC 3′ (SEQ. ID. NO.:7)) which encompassed the deletion and 5′ and 3′ flanking primers whichencompassed the KpnI site and middle T tag on the 5′ end. The final PCRproduct was digested with KpnI and SmaI and ligated into the pS2neo fulllength Akt1 KpnI/SmaI cut vector, effectively replacing the 5′ end ofthe clone with the deleted version.

Human Akt3 gene was amplified by PCR of adult brain cDNA (Clontech)using the amino terminal oligo primer: 5′GASITCAGATCTACCATGAGCGATGTTACCATTGTG 3′ (SEQ. ID. NO.: 8); and thecarboxy terminal oligo primer: 5′ TCTAGATCTTATTCTCGTCCACTTGCAGAG 3′(SEQ.ID. NO.: 9).

These primers included a 5′ EcoRI/BglII site and a 3′ XbaI/BglII sitefor cloning purposes. The resultant PCR product was cloned into theEcoRI and XbaI sites of pGEM4Z (Promega). For expression/purificationpurposes, a middle T tag was added to the 5′ end of the full length Akt3clone using the PCR primer:5′GGTACCATGGAATACATGCCGATGGAAAGCGATGTTACCATTGTGAAG 3′(SEQ. ID. NO.: 10).The resultant PCR product encompassed a 5′ KpnI site which allowed inframe cloning with the biotin tag containing insect cell expressionvector, pS2neo.

Human Akt2 gene was amplified by PCR from human thymus cDNA (Clontech)using the amino terminal oligo primer: 5′AAGCTTAGATCTACCATGAATGAGGTGTCTGTC 3′ (SEQ. ID. NO.: 11); and the carboxyterminal oligo primer: 5′GAATTCGGATCCTCACTCGCGGATGCTGGC 3′ (SEQ. ID.NO.: 12). These primers included a 5′ HindIII/BglII site and a 3′EcoRI/BamHI site for cloning purposes. The resultant PCR product wassubcloned into the HindIII/EcoRI sites of pGem3Z (Promega). Forexpression/purification purposes, a middle T tag was added to the 5′ endof the full length Akt2 using the PCR primer:5′GGTACCATGGAATACATGCCGATGGAAAATGAGGTGTCTGTCATCAAAG 3′ (SEQ. ID. NO.:13). The resultant PCR product was subcloned into the pS2neo vector asdescribed above.

Example 2 Expression of Human Akt Isoforms and Delta-PH-Akt1

The DNA containing the cloned Alai, Akt2, Akt3 and delta-PH-Akt1 genesin the pS2neo expression vector was purified and used to transfectDrosophila S2 cells (ATCC) by the calcium phosphate method. Pools ofantibiotic (G418, 500 μg/mL) resistant cells were selected. Cell wereexpanded to a 1.0 L volume (˜7.0×10⁶/mL), biotin and CuSO₄ were added toa final concentration of 50 μM and 50 mM respectively. Cells were grownfor 72 h at 27° C. and harvested by centrifugation. The cell paste wasfrozen at −70° C. until needed.

Example 3 Purification of Human Akt Isoforms and Delta-PH-Akt1

Cell paste from one liter of S2 cells, described in Example 2, was lysedby sonication with 50 mLs 1% CHAPS in buffer A: (50 mM Tris pH 7.4, 1 mMEDTA, 1 mM EGTA, 0.2 mM AEBSF, 10 μg/mL benzamidine, 5 μg/mL ofleupeptin, aprotinin and pepstatin each, 10% glycerol and 1 mM DTT). Thesoluble fraction was purified on a Protein G Sepharose fast flow(Pharmacia) column loaded with 9 mg/mL anti-middle T monoclonal antibodyand eluted with 75 μM EYMPME (SEQ. ID. NO.: 14) peptide in buffer Acontaining 25% glycerol. Akt/PKB containing fractions were pooled andthe protein purity evaluated by SDS-PAGE. The purified protein wasquantitated using a standard Bradford protocol. Purified protein wasflash frozen on liquid nitrogen and stored at −70° C.

Akt and Akt pleckstrin homology domain deletions purified from S2 cellsrequired activation. Akt and Akt pleckstrin homology domain deletionswere activated (Alessi et al. Current Biology 7:261-269) in a reactioncontaining 10 nM PDK1 (Upstate Biotechnology, Inc.), lipid vesicles (10μM phosphatidylinositol-3,4,5-trisphosphate—Metreya, Inc, 100 μMphosphatidylcholine and 100 μM phosphatidylserine—Avanti Polar lipids,Inc) and activation buffer (50 mM Tris pH7.4, 1.0 mM DTT, 0.1 mM EGTA,1.0 μM Microcystin—LR, 0.1 mM ATP, 10 mM MgCl₂, 333 μg/mL BSA and 0.1 mMEDTA). The reaction was incubated at 22° C. for 4 hours. Aliquots wereflash frozen in liquid nitrogen.

Example 4 Akt Kinase Assays

Activated Akt isoforms and pleckstrin homology domain deletionconstructs were assayed utilizing a GSK-derived biotinylated peptidesubstrate. The extent of peptide phosphorylation was determined byHomogeneous Time Resolved Fluorescence (HTRF) using a lanthanidechelate(Lance)-coupled monoclonal antibody specific for thephosphopeptide in combination with a streptavidin-linked allophycocyanin(SA-APC) fluorophore which will bind to the biotin moiety on thepeptide. When the Lance and APC are in proximity (i.e. bound to the samephosphopeptide molecule), a non-radiative energy transfer takes placefrom the Lance to the APC, followed by emission of light from APC at 665nm.

Materials Required for the Assay:

A. Activated Akt isozyme or pleckstrin homology domain deleted constructB. Akt peptide substrate: GSK3α (S21) Peptide #3928 biotin-GGRARTSSFAEPG(SEQ. ID. NO.:15), Macromolecular Resources.C. Lance labeled anti-phospho GSK3α monoclonal antibody (Cell SignalingTechnology, clone #27).D. SA-APC (Prozyme catalog no. MS lot #896067).

E. Microfluor®B U Bottom Microtiter Plates (Dynex Technologies, Catalogno. 7205). F. Discovery® HTRF Microplate Analyzer, Packard InstrumentCompany.

G. 100× Protease Inhibitor Cocktail (PIC): 1 mg/mL benzamidine, 0.5mg/mL pepstatin, 0.5 mg/mL leupeptin, 0.5 mg/mL aprotinin.H. 10× Assay Buffer: 500 mM HEPES, pH 7.5, 1% PEG, mM EDTA, 1 mM EGTA,1% BSA, 20 mM ∂-Glycerol phosphate.I. Quench Buffer: 50 mM HEPES pH 7.3, 16.6 mM EDTA, 0.1% BSA, 0.1%Triton X-100, 0.17 nM Lance labeled monoclonal antibody clone #27,0.0067 mg/mL SA-APCJ. ATP/MgCl₂ working solution: 1× Assay buffer, 1 mM DTT, 1×PIC, 125 mMKCl, 5% Glycerol, 25 mM MgCl₂, 375 μM ATPK. Enzyme working solution: 1× Assay buffer, 1 mM DTT, 1×PIC, 5%Glycerol, active Akt. The final enzyme concentrations were selected sothat the assay was in a linear response range.L. Peptide working solution: 1× Assay buffer, 1 mM DTT, 1×PIC, 5%Glycerol, 2 μM GSK3 biotinylated peptide #3928

The reaction is assembled by adding 16 pt of the ATP/MgCl₂ workingsolution to the appropriate wells of a 96-well microtiter plate.Inhibitor or vehicle (1.0 μL) is added followed by 10 μL of peptideworking solution. The reaction is started by adding 13 μl, of the enzymeworking solution and mixing. The reaction is allowed to proceed for 50min and then stopped by the addition of 60 μL, HTRF quench buffer. Thestopped reactions were incubated at room temperature for at least 30 minand then read on the Discovery instrument.

IC₅₀ of example compounds to Akt1 kinase and Akt2 kinase are shown inthe table below.

IC₅₀ (nM) Compound Akt1 Akt2 EXAMPLE 2-10 6.0 44.4 EXAMPLE 2-14 0.3 4.0EXAMPLE 3-7 0.4 4.0 EXAMPLE 3-14 0.3 11 EXAMPLE 3-20 0.5 6.3 EXAMPLE5-16 4.7 84.3

Procedure for Streptavidin Flash Plate Assay: Step 1:

A 1 μL solution of the test compound in 100% DMSO was added to 20 μL of2× substrate solution (20 μM GSK3 Peptide, 300 μM ATP, 20 mM MgCl₂, 20μCi/mL[γ³³P] ATP, 1× Assay Buffer, 5% glycerol, 1 mM DTT, 1×PIC, 0.1%BSA and 100 mM KCl). Phosphorylation reactions were initiated by adding19 μL of 2× Enzyme solution (6.4 nM active Akt/PKB, 1× Assay Buffer, 5%glycerol, 1 mM DTT, 1×PIC and 0.1% BSA). The reactions were thenincubated at room temperature for 45 minutes.

Step 2:

The reaction was stopped by adding 170 μL of 125 mM EDTA. 200 μL ofstopped reaction was transferred to a Streptavidin Flashplate® PLUS (NENLife Sciences, catalog no. SMP103). The plate was incubated for ≧10minutes at room temperature on a plate shaker. The contents of each wellwas aspirated, and the wells rinsed 2 times with 200 μL TBS per well.The wells were then washed 3 times for 5 minutes with 200 μL TBS perwell with the plates incubated at room temperature on a platform shakerduring wash steps.

The plates were covered with sealing tape and counted using the PackardTopCount with the appropriate settings for counting [³³P] inFlashplates.

Procedure for Streptavidin Filter Plate Assay: Step 1:

The enzymatic reactions as described in Step 1 of the Streptavidin FlashPlate Assay above were performed.

Step 2:

The reaction was stopped by adding 20 μL of 7.5M GuanidineHydrochloride. 50 μL of the stopped reaction was transferred to theStreptavidin filter plate (SAM²™ Biotin Capture Plate, Promega, catalogno. V7542) and the reaction was incubated on the filter for 1-2 minutesbefore applying vacuum.

The plate was then washed using a vacuum manifold as follows: 1) 4×200μL/well of 2M NaCl; 2) 6×200 μL/well of 2M NaCl with 1% H₃PO₄; 3) 2×200μL/well of diH₂O; and 4) 2×100 pt/well of 95% Ethanol. The membraneswere then allowed to air dry completely before adding scintillant.

The bottom of the plate was sealed with white backing tape, 30 pt/wellof Microscint 20 (Packard Instruments, catalog no. 6013621) was added.The top of the plate was sealed with clear sealing tape, and the platethen counted using the Packard TopCount with the appropriate settingsfor [³³P] with liquid scintillant.

Procedure for Phosphocellulose Filter Plate Assay: Step 1:

The enzymatic reactions were performed as described in Step 1 of theStreptavidin Flash Plate Assay (above) utilizing KKGGRARTSSFAEPG (SEQ.ID. NO.: 16) as the substrate in place of biotin-GGRARTSSFAEPG.

Step 2:

The reaction was stopped by adding 20 μL of 0.75% H₃PO₄. 50 μL ofstopped reaction was transferred to the filter plate (UNIFILTER™,Whatman P81 Strong Cation Exchanger, White Polystyrene 96 Well Plates,Polyfiltronics, catalog no. 7700-3312) and the reaction incubated on thefilter for 1-2 minutes before applying vacuum.

The plate was then washed using a vacuum manifold as follows: 1) 9×200μL/well of 0.75% H₃PO₄; and 2) 2×200 μL/well of diH₂0. The bottom of theplate was sealed with white backing tape, then 30 pt/well of Microscint20 was added. The top of the plate was sealed with clear sealing tape,and the plate counted using the Packard TopCount with the appropriatesettings for [³³P] and liquid scintillant.

PKA Assay:

Each individual PKA assay consists of the following components:A. 5×PKA assay buffer (200 mM Tris pH7.5, 100 mM MgCl₂, 5 mM∂-mercaptoethanol, 0.5 mM EDTA)B. 50 μM stock of Kemptide (Sigma) diluted in waterC. ³³P-ATP prepared by diluting 1.0 μL ³³P-ATP [10 mCi/mL] into 200 μLof a 50 μM stock of unlabeled ATPD. 10 μL of a 70 nM stock of PKA catalytic subunit (UBI catalog #14-114)diluted in 0.5 mg/mL BSAE. PKA/Kemptide working solution: equal volumes of 5×PKA assay buffer,Kemptide solution and PKA catalytic subunit.

The reaction is assembled in a 96 deep-well assay plate. The inhibitoror vehicle (10 μL) is added to 10 μL of the ³³P-ATP solution. Thereaction is initiated by adding 30 μL of the PKA/Kemptide workingsolution to each well. The reactions were mixed and incubated at roomtemperature for 20 min. The reactions were stopped by adding 50 μL of100 mM EDTA and 100 mM sodium pyrophosphate and mixing.

The enzyme reaction product (phosphorylated Kemptide) was collected onp81 phosphocellulose 96 well filter plates (Millipore). To prepare theplate, each well of a p81 filter plate was filled with 75 mM phosphoricacid. The wells were emptied through the filter by applying a vacuum tothe bottom of the plate. Phosphoric acid (75 mM, 170 μL) was added toeach well. A 30 μL aliquot from each stopped PKA reaction was added tocorresponding wells on the filter plate containing the phosphoric acid.The peptide was trapped on the filter following the application of avacuum and the filters were washed 5 times with 75 mM phosphoric acid.After the final wash, the filters were allowed to air dry. Scintillationfluid (30 μL) was added to each well and the filters counted on aTopCount (Packard).

PKC Assay:

Each PKC assay consists of the following components:A. 10×PKC co-activation buffer: 2.5 mM EGTA, 4 mM CaCl₂B. 5×PKC activation buffer: 1.6 mg/mL phosphatidylserine, 0.16 mg/mLdiacylglycerol, 100 mM Tris pH 7.5, 50 mM MgCl₂, 5 mM ∂-mercaptoethanolC. ³³P-ATP prepared by diluting 1.0 μL ³³P-ATP [10 mCi/mL] into 100 μLof a 100 μM stock of unlabeled ATPD. Myelin basic protein (350 μg/mL, UBI) diluted in waterE. PKC (50 ng/mL, UBI catalog #14-115) diluted into 0.5 mg/mL BSAF. PKC/Myelin Basic Protein working solution: Prepared by mixing 5volumes each of PKC co-activation buffer and Myelin Basic protein with10 volumes each of PKC activation buffer and PKC.

The assays were assembled in 96 deep-well assay plates. Inhibitor orvehicle (10 μL) was added to 5.0 μl, of ³³P-ATP. Reactions wereinitiated with the addition of the PKC/Myelin Basic Protein workingsolution and mixing. Reactions were incubated at 30° C. for 20 min. Thereactions were stopped by adding 50 μL of 100 mM EDTA and 100 mM sodiumpyrophosphate and mixing. Phosphorylated Myelin Basic Protein wascollected on PVDF membranes in 96 well filter plates and quantitated byscintillation counting.

Compounds of the instant invention described in the Schemes and Tableswere tested in the assay described above and were found to have IC₅₀ of≦50 μM against one or more of Akt1, Akt2 and Akt3.

Example 5 Cell Based Assays to Determine Inhibition of Akt/PKB

Cells (for example LnCaP or a PTEN(−/−) tumor cell line with activatedAkt/PKB) were plated in 100 mm dishes. When the cells were approximately70 to 80% confluent, the cells were refed with 5 mLs of fresh media andthe test compound added in solution. Controls included untreated cells,vehicle treated cells and cells treated with either LY294002 (Sigma) orwortmanin (Sigma) at 20 μM or 200 nM, respectively. The cells wereincubated for 2, 4 or 6 hrs, and the media removed, the cells werewashed with PBS, scraped and transferred to a centrifuge tube. They werepelleted and washed again with PBS. Finally, the cell pellet wasresuspended in lysis buffer (20 mM Tris pH8, 140 mM NaCl, 2 mM EDTA, 1%Triton X-100, 1 mM Na Pyrophosphate, 10 mM beta-Glycerol Phosphate, 10mM NaF, 0.5 mm NaVO₄, 1 μM Microsystine, and 1× Protease InhibitorCocktail), placed on ice for 15 minutes and gently vortexed to lyse thecells. The lysate was spun in a Beckman tabletop ultra centrifuge at100,000×g at 4° C. for 20 min. The supernatant protein was quantitatedby a standard Bradford protocol (BioRad) and stored at −70° C. untilneeded.

Proteins were immunoprecipitated (IP) from cleared lysates as follows:For Akt1/PKBI, lysates are mixed with Santa Cruz se-7126 (D-17) in NETN(100 mM NaCl, 20 mM Tris pH 8.0, 1 mM EDTA, 0.5% NP-40) and Protein A/GAgarose (Santa Cruz se-2003) was added. For Akt2/PKB∂, lysates weremixed in NETN with anti-Akt2 agarose (Upstate Biotechnology #16-174) andfor Akt3/PKBK, lysates were mixed in NETN with anti-Akt3 agarose(Upstate Biotechnology #16-175). The IPs were incubated overnight at 4°C., washed and separated by SDS-PAGE.

Western blots were used to analyze total Akt, pThr308 Akt1, pSer473Akt1, and corresponding phosphorylation sites on Akt2 and Akt3, anddownstream targets of Akt using specific antibodies (Cell SignalingTechnology): Anti-Total Akt (cat. no. 9272), Anti-Phospho Akt Serine 473(cat. no. 9271), and Anti-Phospho Akt Threonine 308 (cat. no. 9275).After incubating with the appropriate primary antibody diluted inPBS+0.5% non-fat dry milk (NFDM) at 4° C. overnight, blots were washed,incubated with Horseradish peroxidase (HRP)-tagged secondary antibody inPBS+0.5% NFDM for 1 hour at room temperature. Proteins were detectedwith ECL Reagents (Amersham/Pharmacia Biotech RPN2134).

Example 6 Heregulin Stimulated Akt Activation

MCF7 cells (a human breast cancer line that is PTEN^(+/+)) were platedat 1×10⁶ cells per 100 mM plate. When the cells were 70-80% confluent,they were refed with 5 mL of serum free media and incubated overnight.The following morning, compound was added and the cells were incubatedfor 1-2 hrs, after which time heregulin was added (to induce theactivation of Akt) for 30 minutes and the cells were analyzed asdescribed above.

Example 7 Inhibition of Tumor Growth

In vitro/in vivo efficacy of an inhibitor of the growth of cancer cellsmay be confirmed by several protocols well known in the art.

In vitro, 2000-6000 cells/well are seeded into triplicate wells in 96well plate in complete medium (RPMI-1640 supplemented with 10%heat-inactivated fetal bovine serum (FBS)) and incubated at 37° C./5%CO₂ overnight. The next day, inhibitors are added as a dilution seriesin complete medium (final DMSO concentration in the assay is 0.1%). Theplates are incubated at 37° C./5% CO₂ for 72-96 hours. The number ofviable cells is then measured using the CellTiter-Glo kit (Promega). Theluminescence signals are measured using ARVO/Victor 3 plate reader(Perkin-Elmer). The data are fitted with a four parameter dose-responseequation and the inflection point of the least square fit curve orconcentration at 50% inhibition is determined as an IC₅₀ value.

In vivo, human tumor cell lines which exhibit a deregulation of the PI3Kpathway (such as LnCaP, PC3, C33a, OVCAR-3, MDA-MB-468, A2780 or thelike) are injected subcutaneously into the left flank of 6-10 week oldfemale nude (also male mice [age 10-14 weeks] are used for prostatetumor xenografts [LnCaP and PC3]) mice (Harlan), or female nude rats(F344/N Jcl-rnu) (CLEA Japan) on day 0. The mice or rats are randomlyassigned to a vehicle, compound or combination treatment group. Daily orevery other day subcutaneous or oral administration begins on day 1 andcontinues for the duration of the experiment. Alternatively, theinhibitor test compound may be administered by a continuous infusionpump. Compound, compound combination or vehicle is delivered in a totalvolume of 0.2 mL. Tumors are excised and weighed when all of thevehicle-treated animals exhibited lesions of 0.5-1.0 cm in diameter,typically 4 to 5.5 weeks after the cells were injected. The averageweight of the tumors in each treatment group for each cell line iscalculated.

Example 8 Spot Multiplex Assay

This procedure describes a sandwich immunoassay used to detect multiplephosphorylated proteins in the same well of a 96 well format plate. Celllysates are incubated in 96-well plates on which different captureantibodies are placed on spatially distinct spots in the same well.Phosphorylation-specific rabbit polyclonal antibodies are added and thecomplex is detected by an anti-rabbit antibody labeled with anelectrochemiluminescent tag.

96-Well LNCaP Plates+/−Compounds:

Spin in Beckman J6 1200 rpm 10 min, aspirate media. Add 500/well: TBS(Pierce #28376-20 mM Tris pH 7.5, 150 mM NaCl)+1% Triton X-100+Proteaseand Phosphatase Inhibitors. Wrap in plastic wrap, place in −70° C.freezer until completely frozen. Block Multiplex Plates (Meso ScaleDiscovery, Gaithersburg, Md.) with 3% Blocker A in 1×Tris Wash Buffer,150 μl/well. Cover with plate sealer, incubate on Micromix shaker RT 2 h(minimum). Wash with 1×RCM 51 (TTBS). Thaw cell lysate plates on ice,add 40 μl lysate/well into blocked plates. Cover with plate sealer,incubate on Micromix shaker 4° C., O/N, Wash with 1×RCM 51. DiluteSecondary Antibodies in 1% Blocker A in 1× Tris Wash Buffer: Antiphospho AKT (T308), Anti phospho Tuberin (T1462), alone or incombination. Add 25 μl/well, cover with plate sealer, incubate onMicromix shaker RT 3 h. Wash with 1×RCM 51. Dilute Ru-GAR in 1% BlockerA in 1× Tris Wash Buffer. Add 25 μl/well, cover with plate sealer,incubate on Micromix shaker RT 1 h. Wash with 1×RCM 51. Dilute 4× ReadBuffer T to 1× with Water, add 200 μl diluted Read Buffer/well Read onSector 6000 Imager.

Protease and Phosphatase Inhibitors:

Microcystin-LR, Calbiochem #475815 to 1 μM final concentration(stock-500 μM)

Calbiochem #524624, 100× Set I Calbiochem #524625, 100× Set IICalbiochem #539134, 100× Set III Anti Phospho AKT (T308): Cell SignalingTechnologies #9275 Anti Phospho Tuberin (T1462): Covance AffinityPurified (Rabbits MS 2731/2732)

Ru-GAR=Ruthenylated Goat anti Rabbit

10× Tris Wash Buffer, Blocker A and 4× Read Buffer T 10×RCM 51 (10×TTBS,RCM 51) 1×=20 mM Tris pH 7.5, 140 mM NaCl, 0.1% Tween-20 Example 9Cell-Based (In-Vivo) Assay

This procedure describes a cell-based (in vivo) activity assay for theAkt serine/threonine kinase. Activated endogenous Akt is capable ofphosphorylating specific Akt substrate (GSK3β) peptide which isbiotinylated. Detection is performed by Homogeneous Time ResolvedFluorescence (HTRF) using a Europium Kryptate [Eu(K)] coupled antibodyspecific for the phosphopeptide and streptavidin linked XL665fluorophore which will bind to the biotin moiety on the peptide. Whenthe [Eu(K)] and XL665 are in proximity (i.e. bound to the samephosphopeptide molecule) a non-radiative energy transfer takes placefrom the Eu(K) to the XL665, followed by emission of light from XL665 at665 nm.

The assay can be used to detect inhibitors of all three Akt isozymes(Akt1, Akt2, and Akt3) from multiple different species if specificantibodies to each exist.

Materials and Reagents

A. Cell Culture Microtiter Flat Bottom 96 well plates, Corning Costar,Catalog no. 3598B. Reacti-Bind Protein A Coated 96-well plates, Pierce, Catalog no15130.C. Reacti-Bind Protein G Coated 96-well plates, Pierce, Catalog no15131.

D. Micromix 5 Shaker. E. Microfluor®B U Bottom Microtiter Plates, DynexTechnologies, Catalog no. 7205.

F. 96 Well Plate Washer, Bio-Tek Instruments, Catalog no. EL 404.

G. Discovery® HTRF Microplate Analyzer, Packard Instrument Company.Buffer Solutions

A. IP Kinase Cell Lysis Buffer: 1×TBS; 0.2% Tween 20; 1× ProteaseInhibitor Cocktail III (Stock is 100×, Calbiochem, 539134); 1×Phosphatase Inhibitor Cocktail I (Stock is 100×, Calbiochem, 524624);and 1× Phosphatase Inhibitor Cocktail II (Stock is 100×, Calbiochem,524625).B. 10× Assay Buffer: 500 mM Hepes pH 7.5; 1% PEG; 1 mM EDTA; 1 mM EGTA;and 20 mM β-glycerophosphate.

C. IP Kinase Assay Buffer: 1× Assay Buffer; 50 mM KCl; 150 μM ATP; 10 mMMgCl₂; 5% Glycerol; 1 mM DTT; 1 Tablet Protease Inhibitor Cocktail per50 mL Assay Buffer; and 0.1% BSA

D. GSK3β Substrate Solution: IP Kinase Assay Buffer; and 500 nMBiotinylated GSK30 peptide.

E. Lance Buffer: 50 mM Hepes pH 7.5; 0.1% BSA; and 0.1% Triton X-100. F.Lance Stop Buffer: Lance Buffer; and 33.3 mM EDTA.

G. Lance Detection Buffer: Lance Buffer; 13.3 μg/mL SA-APC; and 0.665 nMEuK Ab a-phospho (Ser-21) GSK3β

Multi-Step Immunoprecipitation Akt Kinase Assay Day 1

A. Seed C33a cells Step: Plate 60,000 C33a cells/well in 96 wellmicroliter plate.B. Incubate cells overnight at 37° C.

Day 2

D. Compound Addition Step: Add compounds in fresh media (alpha-MEM/10%FBS, room temp) to 96 well plate from above and incubate for 5 hrs intissue culture incubator.E. Cell Lysis Step: Aspirate media and add 100 μl of IP Kinase CellLysis Buffer.F. Freeze 96 well microtiter plate at −70° C. (NOTE: This step can bedone for a minimum of 1 hour or overnight.)

Day 3

G. Coat Protein A/G 96 well plate Step: Add appropriate concentration ofα-Akt antibody (Akt1, Akt2, or Akt3) in a 100 μl of PBS to the followingwells:

α-Akt 1 (20 ng/well) B2 thru B10 α-Akt 2 (rabbit-human, dog) (50ng/well) B2 thru B10 α-Akt 2 (sheep-mouse, rat) (100 ng/well) B2 thruB10 α-Akt 3 (20 ng/well/100 ul) B2 thru B10

Control-IgG: B11-G11 on every plate

AKT1: rabbit IgG 20 ng/well in 100 μl PBS, Santa Cruz se-2027

AKT2 (for human tumor and dog tissues) rabbit IgG 50 ng/well in 100 μLPBS

AKT2 (for rats and mice) sheep IgG 100 ng/well in 100 μL PBS, Santa Cruzsc-2717

AKT3 rabbit IgG 20 ng/well in 100 μL PBS

H. Incubate in the cold room (+4° C.) for 4 hours on the Micromix 5(Form 20; Attitude 2) (NOTE: Attitude depends on which Micromix 5machine).I. Aspirate off α-Akt antibody solution and add 100 μl of PBS to eachwell.J. Akt Immunoprecipitation Step: To the 100 μl of PBS from Step(I) add 5μl of thawed cell lysate for Akt1 plates and 10 μl of thawed cell lysatefor Akt2 plates. NOTE: Thaw cell lysate on ice. Mix thawed lysate bypipetting up & down 10× before transferring to antibody plates. Keep thecell lysate plates on ice. After transfer of cell lysate to the antibodyplates refreeze the cell lysate plates at −70° C.K. Incubate in the cold room (+4° C.) overnight on Micromix 5 shaker(form 20, attitude 3).

Day 4

L. Immunoprecipitation Plate Wash Step: Wash 96 well plates 1× with TTBS(RCM 51, 1×=2 cycles) using the 96-Well Plate Washer. Fill wells withTTBS and incubate for 10 minutes. Wash 96 well plates 2× with TTBS.(NOTE: Prime plate washer before use: 1. Check buffer reservoirs, makingsure they are full and 2. empty waste containers.M. Manual Plate Wash Step: Add 180 μl of IP Kinase Assay buffer.N. Start Akt Enzyme Reaction: Aspirate supernatant. Add 60 μl of GSK3βSubstrate Solution.O. Incubate for 2.5 hours on Micromix 5 shaker @ RT. NOTE: Time ofincubation should be adjusted so that the ratio of Column 10/Column 11is not >10.P. Combine 30 μl of Lance Detection Buffer with 30 μl of Lance StopBuffer (60 μl total/well) and add to Microfluor U bottom 96 well blackplates.Q. Stop Akt Enzyme Reaction: Transfer 40 μl of Akt Enzyme Reaction Mixfrom Protein A/G 96 well plate from Step (O) to Microfluor U bottom 96well black plates from Step (P).U. Incubate at room temperature for 1-2 hrs on Micromix 5 shaker (form20, attitude 3), then read with the Discovery HTRF Microplate Analyzerusing Akt program.

IP Kinase Cell Lysis Buffer 1×TBS 0.25% Tween 20 (Fisher BP337-500) 1×Protease Inhibitor Cocktail III (Stock is 100×, Calbiochem, 539134) 1×Phosphatase Inhibitor Cocktail I (Stock is 100×, Calbiochem, 524624) 1×Phosphatase Inhibitor Cocktail II (Stock is 100×, Calbiochem, 524625) 1μM Microcystin LR (Calbiochem 475815) IP Kinase Assay Buffer 50 mM HepespH 7.5 0.1% PEG (Sigma P-3265) 0.1 mM EDTA (USB 15694) 0.1 mM EGTA(Sigma E8145-50G)

2 mM β-glycerophosphate (Sigma G-6376)50 mM KCl (Fisher P-217) (1M stock, RT)

150 μM ATP (Sigma) 10 mM MgCl₂ (Sigma M-1028) 5% Glycerol (FisherG33-500) 1 mM DTT (Sigma D0632-25G) 1 Tablet Protease Inhibitor Cocktailper 50 mL (Roche 11 836 145 001) 0.1% BSA (Roche 03 117 405 001) GSK3βSubstrate Solution IP Kinase Assay Buffer

500 nM Biotinylated GSK3β peptide (Biotin-GGRARTSSFAEPG-COOH)

Lance Stop Buffer 25 in M Hepes pH 7.5 0.05% BSA 0.05% Triton X-100 16.7mM EDTA Lance Detection Buffer

6.65 μg/mL SA-APC (Perkin Elmer CR130-100)0.665 nM EuK Ab a-phospho (Ser-21) GSK3β monoclonal antibody in LanceStop Buffer

1. A compound according to Formula A:

wherein: E, F, G, H, I, J, K, L and M are independently selected from: Cor N, wherein each E, F, G, H, I, J, K, L and M is optionallysubstituted with R¹; a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; p isindependently 0, 1, 2, 3, 4 or 5; Ring Y is (C₄-C₇)cycloalkyl, saidcycloalkyl is substituted with R^(x), and further optionally substitutedwith one or more substituents selected from: (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, CO₂H, halo, CN, OH and NR⁷R⁸, saidalkyl, cycloalkyl and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸; Ring Z is selectedfrom: (C₃-C₈)cycloalkyl, aryl, heteroaryl and heterocyclyl; R¹ isselected from: H, oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)-aryl,(C═O)_(a)O_(b)(C₂-C₁₀)alkenyl, (C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H,halo, OH, O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)_(m)NR⁷R⁸, SH,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶; R² isindependently selected from: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, SH, S(O)_(m)NR⁷R⁸,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶; R⁶ is:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo, CN, OH,O_(b)(C₁-C₆)perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO, (N═O)R⁷R⁸,S(O)_(m)NR⁷R⁸, SH, S(O)_(m)—(C₁-C₁₀)alkyl or(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, said alkyl, aryl, alkenyl, alkynyl,heterocyclyl, and cycloalkyl optionally substituted with one or moresubstituents selected from R^(6a); R^(6a) is selected from:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, O_(a)(C₁-C₃)perfluoroalkyl,(C₀-C₆)alkylene-S(O)_(m)R^(a), SH, oxo, OH, halo, CN, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl, (C₀-C₆)alkylene-aryl,(C₀-C₆)alkylene-heterocyclyl, (C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a),(C₀-C₆)alkylene-CO₂R^(a), C(O)H, and (C₀-C₆)alkylene-CO₂H, said alkyl,alkenyl, alkynyl, cycloalkyl, aryl, and heterocyclyl is optionallysubstituted with up to three substituents selected from R^(b), OH,(C₁-C₆)alkoxy, halogen, CO₂H, CN, O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, andN(R^(b))₂; R⁷ and R⁸ are independently selected from: H,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)-heterocyclyl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, SH, SO₂R^(a), and (C═O)_(a)NR^(b) ₂, said alkyl,cycloalkyl, aryl, heterocyclyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(6a), or R⁷and R⁸ can be taken together with the nitrogen to which they areattached to form a monocyclic or bicyclic heterocycle with 3-7 membersin each ring and optionally containing, in addition to the nitrogen, oneor two additional heteroatoms selected from N, O and S, said monocyclicor bicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a); R^(a) is (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, aryl, or heterocyclyl; R^(b) is independently: H,(C₁-C₆)alkyl, aryl, heterocyclyl, (C₃-C₆)cycloalkyl,(C═O)_(a)O_(b)(C₁-C₆)alkyl, or S(O)_(m)R^(a); and R^(x) is selectedfrom: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸, said alkyl, andalkoxy is optionally substituted with one or more substituents selectedfrom halo, CN, OH and NR⁷R⁸; or a pharmaceutically acceptable salt or astereoisomer thereof.
 2. A compound according to claim 1 of Formula A,wherein:

is selected from:

Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H; and all other substituents andvariables are as defined in claim 1; or a pharmaceutically acceptablesalt or a stereoisomer thereof.
 3. A compound according to claim 2 ofFormula B:

wherein: Ring Y is cyclobutyl, said cyclobutyl is substituted withR^(x), and further optionally substituted with one or more substituentsselected from: (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, CO₂H,halo, CN, OH and NR⁷R⁸, said alkyl, cycloalkyl, and alkoxy is optionallysubstituted with one or more substituents selected from halo, CN, OH andNR⁷R⁸; p is 0, 1 or 2; R² is independently selected from: (C₁-C₆)alkyl,(C₁-C₆)alkoxy, CO₂H, halo, OH and NH₂; and all other substituents andvariables are as defined in claim 2; or a pharmaceutically acceptablesalt or a stereoisomer thereof.
 4. A compound according to claim 1 ofFormula C:

wherein: a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; p is 0, 1 or 2; R² isindependently selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, CO₂H, halo, OHand NH₂; R¹ is selected from: H, oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)_(m)NR⁷R⁸, SH,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶; R⁶ is:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo, CN, OH,O_(b)(C₁-C₆)perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO, (N═O)R⁷R⁸,S(O)_(m)NR⁷R⁸, SH, S(O)_(m)—(C₁-C₁₀)alkyl or(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, said alkyl, aryl, alkenyl, alkynyl,heterocyclyl, and cycloalkyl optionally substituted with one or moresubstituents selected from R^(6a); R^(6a) is selected from:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, O_(a)(C₁-C₃)perfluoroalkyl,(C₀-C₆)alkylene-S(O)_(m)R^(a), SH, oxo, OH, halo, CN, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl, (C₀-C₆)alkylene-aryl,(C₀-C₆)alkylene-heterocyclyl, (C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a),(C₀-C₆)alkylene-CO₂R^(a), C(O)H, and (C₀-C₆)alkylene-CO₂H, said alkyl,alkenyl, alkynyl, cycloalkyl, aryl, and heterocyclyl is optionallysubstituted with up to three substituents selected from R^(b), OH,(C₁-C₆)alkoxy, halogen, CO₂H, CN, O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, andN(R^(b))₂; R⁷ and R⁸ are independently selected from: H,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)-heterocyclyl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, SH, SO₂R^(a), and (C═O)_(a)NR^(b) ₂, said alkyl,cycloalkyl, aryl, heterocyclyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(6a), or R⁷and R⁸ can be taken together with the nitrogen to which they areattached to form a monocyclic or bicyclic heterocycle with 3-7 membersin each ring and optionally containing, in addition to the nitrogen, oneor two additional heteroatoms selected from N, O and S, said monocyclicor bicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a); R^(a) is (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, aryl, or heterocyclyl; and R^(b) is independently: H,(C₁-C₆)alkyl, aryl, heterocyclyl, (C₃-C₆)cycloalkyl,(C═O)_(a)O_(b)(C₁-C₆)alkyl, or S(O)_(m)R^(a); R^(x) is selected from:(C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸, said alkyl, and alkoxyis optionally substituted with one or more substituents selected fromhalo, CN, OH and NR⁷R⁸; R^(y) is selected from: H, (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, halo, CN and OH, said alkyl,cycloalkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸; and Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H; or a pharmaceuticallyacceptable salt or a stereoisomer thereof.
 5. A compound according toclaim 1 of Formula D:

wherein: a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; p is 0, 1 or 2; R² isindependently selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, CO₂H, halo, OHand NH₂; R¹ is selected from: H, oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)_(m)NR⁷R⁸, SH,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶; R⁶ is:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo, CN, OH,O_(b)(C₁-C₆)perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO, (N═O)R⁷R⁸,S(O)_(m)NR⁷R⁸, SH, S(O)_(m)(C₁-C₁₀)alkyl or(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, said alkyl, aryl, alkenyl, alkynyl,heterocyclyl, and cycloalkyl optionally substituted with one or moresubstituents selected from R^(6a); R^(6a) is selected from:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, O_(a)(C₁-C₃)perfluoroalkyl,(C₀-C₆)alkylene-S(O)_(m)R^(a), SH, oxo, OH, halo, CN, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl, (C₀-C₆)alkylene-aryl,(C₀-C₆)alkylene-heterocyclyl, (C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a),(C₀-C₆)alkylene-CO₂R^(a), C(O)H, and (C₀-C₆)alkylene-CO₂H, said alkyl,alkenyl, alkynyl, cycloalkyl, aryl, and heterocyclyl is optionallysubstituted with up to three substituents selected from R^(b), OH,(C₁-C₆)alkoxy, halogen, CO₂H, CN, O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, andN(R^(b))₂; R⁷ and R⁸ are independently selected from: H,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)-heterocyclyl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, SH, SO₂R^(a), and (C═O)_(a)NR^(b) ₂, said alkyl,cycloalkyl, aryl, heterocyclyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(6a), or R⁷and R⁸ can be taken together with the nitrogen to which they areattached to form a monocyclic or bicyclic heterocycle with 3-7 membersin each ring and optionally containing, in addition to the nitrogen, oneor two additional heteroatoms selected from N, O and S, said monocyclicor bicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a); R^(a) is (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, aryl, or heterocyclyl; and R^(b) is independently: H,(C₁-C₆)alkyl, aryl, heterocyclyl, (C₃-C₆)cycloalkyl,(C═O)_(a)O_(b)(C₁-C₆)alkyl, or S(O)_(m)R^(a); R^(x) is selected from:(C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸, said alkyl, and alkoxyis optionally substituted with one or more substituents selected fromhalo, CN, OH and NR⁷R⁸; R^(y) is selected from: H, (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, halo, CN and OH, said alkyl,cycloalkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸; and Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H; or a pharmaceuticallyacceptable salt or a stereoisomer thereof.
 6. A compound according toclaim 1 of Formula E:

wherein: a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; p is 0, 1 or 2; R² isindependently selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, CO₂H, halo, OHand NH₂; R¹ is selected from: H, oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)_(m)NR⁷R⁸, SH,S(O)_(m)—(C₁-C₁₀)alkyl and (C═O)_(a)O_(b)-heterocyclyl, said alkyl,aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionallysubstituted with one or more substituents selected from R⁶; R⁶ is:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)aryl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo, CN, OH,O_(b)(C₁-C₆)perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO, (N═O)R⁷R⁸,S(O)_(m)NR⁷R⁸, SH, S(O)_(m)(C₁-C₁₀)alkyl or(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, said alkyl, aryl, alkenyl, alkynyl,heterocyclyl, and cycloalkyl optionally substituted with one or moresubstituents selected from R^(6a); R^(6a) is selected from:(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, O_(a)(C₁-C₃)perfluoroalkyl,(C₀-C₆)alkylene-S(O)_(m)R^(a), SH, oxo, OH, halo, CN, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl, (C₀-C₆)alkylene-aryl,(C₀-C₆)alkylene-heterocyclyl, (C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a),(C₀-C₆)alkylene-CO₂R^(a), C(O)H, and (C₀-C₆)alkylene-CO₂H, said alkyl,alkenyl, alkynyl, cycloalkyl, aryl, and heterocyclyl is optionallysubstituted with up to three substituents selected from R^(b), OH,(C₁-C₆)alkoxy, halogen, CO₂H, CN, O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, andN(R^(b))₂; R⁷ and R⁸ are independently selected from: H,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)-heterocyclyl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, SH, SO₂R^(a), and (C═O)_(a)NR^(b) ₂, said alkyl,cycloalkyl, aryl, heterocyclyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(6a), or R⁷and R⁸ can be taken together with the nitrogen to which they areattached to form a monocyclic or bicyclic heterocycle with 3-7 membersin each ring and optionally containing, in addition to the nitrogen, oneor two additional heteroatoms selected from N, O and S, said monocyclicor bicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a); R^(a) is (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, aryl, or heterocyclyl; and R^(b) is independently: H,(C₁-C₆)alkyl, aryl, heterocyclyl, (C₃-C₆)cycloalkyl,(C═O)_(a)O_(b)(C₁-C₆)alkyl, or S(O)_(m)R^(a); R^(x) is selected from:(C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸, said alkyl, and alkoxyis optionally substituted with one or more substituents selected fromhalo, CN, OH and NR⁷R⁸; R^(y) is selected from: H, (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, halo, CN and OH, said alkyl,cycloalkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸; and Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H; or a pharmaceuticallyacceptable salt or a stereoisomer thereof.
 7. A compound according toclaim 1 of Formula C-1:

wherein: R¹ is imidazolyl, triazolyl, or pyrimidyl, said imidazolyl,triazolyl, and pyrimidyl is optionally substituted with one or more(C₁-C₆)alkyl; R⁷ and R⁸ are independently selected from: H, and(C₁-C₆)alkyl; R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo,OH and NR⁷R⁸, said alkyl, and alkoxy is optionally substituted with oneor more substituents selected from halo, CN, OH and NR⁷R⁸; R^(y) isselected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, halo,CN and OH, said alkyl, cycloalkyl, and alkoxy is optionally substitutedwith one or more substituents selected from halo, CN, OH and NR⁷R⁸; andBond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H; or a pharmaceuticallyacceptable salt thereof.
 8. A compound according to claim 1 of FormulaD-1:

wherein: R¹ is pyridyl, said pyridyl is optionally substituted with oneor more (C₁-C₆)alkyl; R⁷ and R⁸ are independently selected from: H, and(C₁-C₆)alkyl; R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo,OH and NR⁷R⁸, said alkyl, and alkoxy is optionally substituted with oneor more substituents selected from halo, CN, OH and NR⁷R⁸; R^(y) isselected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, halo,CN and OH, said alkyl, cycloalkyl, and alkoxy is optionally substitutedwith one or more substituents selected from halo, CN, OH and NR⁷R⁸; andBond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H; or a pharmaceuticallyacceptable salt or a stereoisomer thereof.
 9. A compound according toclaim 1 of Formula D-1:

wherein: R¹ is (C₁-C₆)alkyl, NR⁷R⁸, phenyl, pyridyl, pyrimidyl,pyrazinyl, azetidinyl, piperidinyl, or morpholinyl, said alkyl, phenyl,pyridyl, pyrimidyl, pyrazinyl, azetidinyl, piperidinyl, and morpholinylis optionally substituted with one or more substituents selected from:(C₁-C₆)alkyl, halo, OH, and NR⁷R⁸, said alkyl is optionally substitutedwith one or more substituents selected from: halo, and OH; R⁷ and R⁸ areindependently selected from: H, and (C₁-C₆)alkyl; R^(x) is selectedfrom: (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo, OH and NR⁷R⁸, said alkyl, andalkoxy is optionally substituted with one or more substituents selectedfrom halo, CN, OH and NR⁷R⁸; R^(y) is selected from: H, (C₁-C₆)alkyl,(C₃-C₆)cycloalkyl, (C₁-C₆)alkoxy, halo, CN and OH, said alkyl,cycloalkyl, and alkoxy is optionally substituted with one or moresubstituents selected from halo, CN, OH and NR⁷R⁸; and Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H; or a pharmaceuticallyacceptable salt or a stereoisomer thereof.
 10. A compound according toclaim 1 of Formula E-1:

wherein: R¹ is pyrimidyl, said pyrimidyl is optionally substituted withone or more (C₁-C₆)alkyl; R⁷ and R⁸ are independently selected from: H,and (C₁-C₆)alkyl; R^(x) is selected from: (C₁-C₆)alkyl, (C₁-C₆)alkoxy,halo, OH and NR⁷R⁸, said alkyl, and alkoxy is optionally substitutedwith one or more substituents selected from halo, CN, OH and NR⁷R⁸;R^(y) is selected from: H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,(C₁-C₆)alkoxy, halo, CN and OH, said alkyl, cycloalkyl, and alkoxy isoptionally substituted with one or more substituents selected from halo,CN, OH and NR⁷R⁸; and Bond:

is a single or double bond, provided that when R¹ is oxo, then said bondis a single bond and adjacent N bears H; or a pharmaceuticallyacceptable salt or a stereoisomer thereof.
 11. A compound which isselected from: (1)cis-3-amino-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(2)trans-3-amino-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(3)cis-3-amino-1-methyl-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(4)trans-3-amino-1-methyl-3-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(5)3,3-dimethoxy-1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(6)trans-3-amino-1-methyl-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(7)trans-3-fluoro-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(8)cis-3-fluoro-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(9)3,3-difluoro-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine);(10)trans-3-amino-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(11)cis-3-amino-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(12)trans-3-methoxy-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(13)trans-3-amino-1-cyclopropyl-3-{4-[9-phenyl-3-(2-pyrimidinyl)-[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(14)cis-3-amino-1-cyclopropyl-3-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(15)3-fluoro-3-methyl-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(16)trans-3-amino-1-methyl-3-{-4-[2-phenyl-9-(3-pyridinyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(17)2,2,2-trifluoro-N-(3-hydroxy-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)acetamide;(18)N-(3-(dimethylamino)-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide;(19)N³,N³-dimethyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)-[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-1,3-cyclobutanediamine;(20)N³-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}-1,3-cyclobutanediamine;(21)N-(3,3-difluoro-1-{-4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide;(22)3-(methylamino)-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(23)N-(3-cyano-3-hydroxy-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutyl)-2,2,2-trifluoroacetamide;(24)3-methyl-1-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(25)3-amino1-(hydroxymethyl)-3-{4-[9-phenyl-3-(2-pyrimidinyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(26)3,3-difluoro-1-{4-[3-(1-methyl-1H-imidazol-4-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl])phenyl}cyclobutanamine;(27)3,3-difluoro-1-{-4-[9-phenyl-3-(trifluoromethyl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(28)3,3-difluoro-1-{-4-[9-phenyl-3-(1H-1,2,3-triazol-4-yl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(29)3,3-difluoro-1-{4-[3-(1H-indazol-3-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(30)3,3-difluoro-1-{4-[3-(5-methyl-1H-1,2,4-triazol-3-yl)-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanamine;(31)8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridine-3-carboxamide;(32)8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-amine;(33)8-[4-(1-amino-3,3-difluorocyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-ol;(34)3-amino-3-{4-[9-phenyl-2-(2-pyrimidinyl)[1,2,4]triazolo[5,1-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(35)trans-3-amino-1-ethyl-3-{4-[9-phenyl-3-(pyrimidin-2-yl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(36)trans-3-amino-1-methyl-3-{4-[3-(pyrimidin-2-yl)-9-(thiophen-2-yl)[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-8-yl]phenyl}cyclobutanol;(37)trans-3-amino-1-methyl-3-{4-[9-phenyl-3-(pyrimidin-2-yl)[1,2,4]triazolo[4′,3′:1,2]pyrido[3,4-b]pyrazin-8-yl]phenyl}cyclobutanol;(38)8-[4-(trans-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl]-9-phenyl[1,2,4]triazolo[3,4-f]-1,6-naphthyridin-3-ol;(39)trans-3-amino-1-cyclopropyl-3-{4-[9-(3,4-difluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(40)trans-3-amino-1-cyclopropyl-3-[4-(2,9-diphenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]cyclobutanol;(41)trans-3-amino-3-{4-[9-(4-chlorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol;(42)3-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol;(43)trans-3-amino-1-cyclopropyl-3-{4-[9-(4-methylphenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(44)trans-3-amino-1-methyl-3-{-4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(45)4-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol;(46)trans-3-amino-1-cyclopropyl-3-{4-[9-(4-fluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(47)trans-3-amino-1-cyclopropyl-3-{4-[9-(3-fluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(48)3-amino-3-{4-[9-(2-aminopyrimidin-5-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol;(49)3-{3-[4-(trans-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol;(50)trans-3-amino-3-{4-[9-(6-aminopyridin-3-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol;(51)trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(52)trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(pyridin-3-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(53)trans-3-amino-1-ethyl-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(54)trans-3-amino-1-cyclopropyl-3-{4-[9-(morpholin-4-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(55)trans-3-amino-3-[4-(9-ethyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-methylcyclobutanol;(56)trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(pyrimidin-5-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(57)trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(piperidin-1-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(58)trans-3-amino-3-{4-[9-(2-aminopyridin-4-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-methylcyclobutanol;(59)trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(pyrazin-2-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(60)(3-amino-3-{-4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutyl)methanol;(61)trans-3-amino-1-cyclopropyl-3-{4-[9-(difluoromethyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(62)trans-3-amino-3-{4-[9-(difluoromethyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-methylcyclobutanol;(63)trans-3-amino-3-{4-[9-(3-chlorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol;(64)trans-3-amino-1-cyclopropyl-3-{4-[9-(2-fluorophenyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(65)cis-3-amino-1-methyl-3-{4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(66)trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(trifluoromethyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(67)trans-3-amino-1-cyclopropyl-3-{4-[9-(3-methylphenyl)-2-phenyl][1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol;(68)cis-3-amino-1-methyl-3-{4-[9-(morpholin-4-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(69)(3-amino-3-{-4-[2-phenyl-9-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutyl)methanol;(70)3-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl}benzonitrile;(71)cis-3-amino-1-methyl-3-{4-[2-phenyl-9-(pyrimidin-5-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(72)2-{3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl]phenol;(73)trans-3-amino-3-[4-(9-tert-butyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol;(74)trans-3-amino-1-cyclopropyl-3-(4-{2-phenyl-9-[3-(trifluoromethyl)phenyl][1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol;(75)trans-3-amino-1-cyclopropyl-3-(4-{9-[4-(methylsulfonyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol;(76)trans-3-amino-1-cyclopropyl-3-{4-[2-phenyl-9-(1H-pyrrolo[2,3-b]pyridin-4-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(77)3,3-difluoro-1-{-4-[2-phenyl-9-(1H-1,2,4-triazol-3-yl)[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanamine;(78)trans-3-amino-1-cyclopropyl-3-{4-[9-(2-hydroxypropan-2-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(79)trans-3-amino-1-cyclopropyl-3-[4-(9-methyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]cyclobutanol;(80)trans-3-amino-1-cyclopropyl-3-{4-[9-(dimethylamino)-2-phenyl-[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(81)trans-3-amino-1-cyclopropyl-3-{4-[9-(methylamino)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(82)trans-3-amino-3-[4-(9-amino-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol;(83)trans-3-amino-3-{4-[9-(azetidin-1-yl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}-1-cyclopropylcyclobutanol;(84)trans-3-amino-3-[4-(9-chloro-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]-1-cyclopropylcyclobutanol;(85)trans-3-amino-1-cyclopropyl-3-{4-[9-(methylsulfanyl)-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl]phenyl}cyclobutanol;(86)trans-3-amino-1-cyclopropyl-3-[4-(9-methoxy-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl)phenyl]cyclobutanol;(87)3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-ol;(88)3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-carboxamide;(89)trans-3-amino-1-cyclopropyl-3-(4-{9-[(dimethylamino)methyl]-2-phenyl-[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-3-yl}phenyl)cyclobutanol;(90)N-({3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-yl}methyl)acetamide;and (91)3-[4-(trans-1-amino-3-cyclopropyl-3-hydroxycyclobutyl)phenyl]-N-ethyl-2-phenyl[1,2,4]triazolo[4′,3′:1,6]pyrido[2,3-b]pyrazin-9-carboxamide;or a pharmaceutically acceptable salt or stereoisomer thereof.
 12. Apharmaceutical composition comprising a pharmaceutical carrier, anddispersed therein, a therapeutically effective amount of a compound ofclaim
 1. 13. (canceled)